Low eructation composition and methods for treating and/or preventing cardiovascular disease in a subject with fish allergy/hypersensitivity

ABSTRACT

In various embodiments, the present invention provides pharmaceutical compositions and methods for treating cardiovascular-related disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/179,623 filed Nov. 2, 2018, which is a continuation of U.S. patentapplication Ser. No. 13/990,316 filed Jan. 23, 2014, which is a NationalStage Entry of International Application No. PCT/US2011/062247 filedNov. 28, 2011, which claims the benefit of U.S. Application No.61/417,691 filed Nov. 29, 2010.

BACKGROUND

Cardiovascular disease is one of the leading causes of death in theUnited States and most European countries. It is estimated that over 70million people in the United States alone suffer from a cardiovasculardisease or disorder including but not limited to high blood pressure,coronary heart disease, dislipidemia, congestive heart failure andstroke. A need exists for improved treatments for cardiovasculardiseases and disorders.

It is further estimated that more than 6 million Americans suffer fromsome form of seafood allergy. Sicherer et al., Journal of Allergy andClinical Immunology, Volume 114, Issue 1, Pages 159-165. Therapiesderived from fish oil have been proven effective at loweringtriglycerides, among other effects. Unfortunately, such products arecontraindicated in subjects that are allergic or hypersensitive to fish.

SUMMARY

In various embodiments, the present invention provides methods oftreating and/or preventing cardiovascular-related diseases and, inparticular, a method of blood lipid therapy comprising administering toa subject in need thereof a pharmaceutical composition comprisingeicosapentaenoic acid or a derivative thereof. In one embodiment, thecomposition contains not more than 10%, by weight, docosahexaenoic acidor derivative thereof, substantially no docosahexaenoic acid orderivative thereof, or no docosahexaenoic acid or derivative thereof. Inanother embodiment, eicosapentaenoic acid ethyl ester comprises at least96%, by weight, of all fatty acids present in the composition; thecomposition contains not more than 4%, by weight, of total fatty acidsother than eicosapentaenoic acid ethyl ester; and/or the compositioncontains about 0.1% to about 0.6% of at least one fatty acid other thaneicosapentaenoic acid ethyl ester and docosahexaenoic acid (orderivative thereof).

In various embodiments, the present invention provides pharmaceuticalcompositions and methods of using such compositions to increase plasma,serum and/or red blood cell (RBC) EPA levels and/or to treat or preventcardiovascular-related diseases. In one embodiment, the subject beingtreated is hypersensitive and/or allergic to seafood, shellfish and/orfish. In other embodiments, ingestion of the pharmaceutical compositioncauses no or reduced eructation as compared to placebo and/or tocommercially available omega-3 fatty acid blends (e.g. Lovaza).

In a related embodiment, ingestion of the pharmaceutical composition asset forth herein does not cause skin rash or causes reduced incidence ofskin rash compared to placebo and/or commercially available omega-3fatty acid blends (e.g. Lovaza).

In one embodiment, the invention provides a pharmaceutical compositioncomprising, consisting of or consisting essentially of at least 95% byweight ethyl eicosapentaenoate (EPA-E), about 0.2% to about 0.5% byweight ethyl octadecatetraenoate (ODTA-E), about 0.05% to about 0.25% byweight ethyl nonaecapentaenoate (NDPA-E), about 0.2% to about 0.45% byweight ethyl arachidonate (AA-E), about 0.3% to about 0.5% by weightethyl eicosatetraenoate (ETA-E), and about 0.05% to about 0.32% ethylheneicosapentaenoate (HPA-E). In another embodiment, the composition ispresent in a capsule shell. In another embodiment, the compositioncontains substantially no or no amount of docosahexaenoic acid (DHA) orderivative thereof such as ethyl-DHA (DHA-E), for example not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight. In a relatedembodiment, the composition is substantially free of or is free ofallergens.

In another embodiment, the invention provides a method of increasingserum, plasma and/or red blood cell (RBC) EPA levels comprisingadministering a composition as described herein to a subject in need ofincreased serum, plasma and/or RBC EPA levels.

In a related embodiment, the subject has a baseline EPA plasma, serumand/or RBC level not greater than about 50 μg/g and upon administeringthe composition to the subject for a period of at least about 6 weeks,the subject exhibits at least a 100%, at least a 150%, at least a 200%,at least a 250%, at least 300%, at least 350% or at least 400% increase(change in EPA level divided by baseline EPA level) in plasma, serumand/or RBC EPA levels compared to baseline or placebo control. In arelated embodiment, the subject has a baseline EPA plasma, serum and/orRBC level not greater than about 50 μg/g. In another embodiment, thesubject is provided with an amount of said composition effective toachieve said increases in EPA levels. In another embodiment, the subjectis provided with about 2 g to about 4 g per day of said composition.

In another embodiment, the invention provides a method of treating acardiovascular-related disease in a subject in need thereof, comprisingadministering a composition as described herein to the subject. In arelated embodiment, the subject has a baseline EPA plasma, serum and/orRBC level not greater than about 50 μg/g and upon administering thecomposition to the subject for a period of at least about 6 weeks, thesubject exhibits at least about a 100%, at least about a 150%, at leastabout a 200%, at least about a 250%, at least about a 300%, at leastabout a 350% or at least about a 400% increase in plasma, serum and/orRBC EPA levels compared to baseline or placebo control. In a relatedembodiment, the subject has a baseline EPA plasma, serum and/or RBClevel not greater than about 50 μg/g. In another embodiment, the subjectis provided with about 2 g to about 4 g per day of said composition.

These and other embodiments of the present invention will be disclosedin further detail herein below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1 and 2 show a comparison of the change in plasma/serum EPA levelsobserved with AMR101 treatment in the current study compared to thoseobserved with different EPA compositions in the JELIS study and byGrimsgaard.

FIGS. 3 and 4 show top line results of the study disclosed herein.

DETAILED DESCRIPTION

While the present invention is capable of being embodied in variousforms, the description below of several embodiments is made with theunderstanding that the present disclosure is to be considered as anexemplification of the invention, and is not intended to limit theinvention to the specific embodiments illustrated. Headings are providedfor convenience only and are not to be construed to limit the inventionin any manner. Embodiments illustrated under any heading may be combinedwith embodiments illustrated under any other heading.

The use of numerical values in the various quantitative values specifiedin this application, unless expressly indicated otherwise, are stated asapproximations as though the minimum and maximum values within thestated ranges were both preceded by the word “about.” Also, thedisclosure of ranges is intended as a continuous range including everyvalue between the minimum and maximum values recited as well as anyranges that can be formed by such values. Also disclosed herein are anyand all ratios (and ranges of any such ratios) that can be formed bydividing a disclosed numeric value into any other disclosed numericvalue. Accordingly, the skilled person will appreciate that many suchratios, ranges, and ranges of ratios can be unambiguously derived fromthe numerical values presented herein and in all instances such ratios,ranges, and ranges of ratios represent various embodiments of thepresent invention.

In one embodiment, the invention provides pharmaceutical compositionscomprising eicosapentaenoic acid or a derivative thereof. In oneembodiment, such compositions comprise eicosapentaenoic acid, or apharmaceutically acceptable ester, derivative, conjugate or saltthereof, or mixtures of any of the foregoing, collectively referred toherein as “EPA.” The term “pharmaceutically acceptable” in the presentcontext means that the substance in question does not produceunacceptable toxicity to the subject or interaction with othercomponents of the composition.

In one embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid. In another embodiment, the EPAcomprises an eicosapentaenoic acid ester. In another embodiment, the EPAcomprises a C₁-C₅ alkyl ester of eicosapentaenoic acid. In anotherembodiment, the EPA comprises eicosapentaenoic acid ethyl ester,eicosapentaenoic acid methyl ester, eicosapentaenoic acid propyl ester,or eicosapentaenoic acid butyl ester. In another embodiment, the EPAcomprises all-cis eicosa-5,8,11,14,17-pentaenoic acid ethyl ester.

In another embodiment, the EPA is in the form of ethyl-EPA, lithium EPA,mono-, di- or triglyceride EPA or any other ester or salt of EPA, or thefree acid form of EPA. The EPA may also be in the form of a2-substituted derivative or other derivative which slows down its rateof oxidation but does not otherwise change its biological action to anysubstantial degree.

In another embodiment, the invention provides a composition comprisingEPA for the treatment of hypertriglyceridemia in a subject or subjectsthat is/are allergic to fish. In one embodiment, a subject is diagnosedas allergic to fish (e.g. by prior history, allergic challenge,shellfish mix skin prick test or fish mix skin prick test) or othermethodology) prior to or after initiation of administration of thecomposition of the invention to said subject or subjects.

In another embodiment, the invention provides a composition comprisingEPA as defined herein for the treatment of hypertriglyceridemia in asubject or subjects that is susceptible to eructation or thatexperiences eructation while on other commercially available omega-3fatty acid therapy (e.g. Lovaza).

In another embodiment, the composition is present in a dosage unit (e.g.a capsule) in an amount of about 50 mg to about 5000 mg, about 75 mg toabout 2500 mg, or about 100 mg to about 1000 mg, for example about 75mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575mg, about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700mg, about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825mg, about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950mg, about 975 mg, about 1000 mg, about 1025 mg, about 1050 mg, about1075 mg, about 1100 mg, about 1025 mg, about 1050 mg, about 1075 mg,about 1200 mg, about 1225 mg, about 1250 mg, about 1275 mg, about 1300mg, about 1325 mg, about 1350 mg, about 1375 mg, about 1400 mg, about1425 mg, about 1450 mg, about 1475 mg, about 1500 mg, about 1525 mg,about 1550 mg, about 1575 mg, about 1600 mg, about 1625 mg, about 1650mg, about 1675 mg, about 1700 mg, about 1725 mg, about 1750 mg, about1775 mg, about 1800 mg, about 1825 mg, about 1850 mg, about 1875 mg,about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg, about 2000mg, about 2025 mg, about 2050 mg, about 2075 mg, about 2100 mg, about2125 mg, about 2150 mg, about 2175 mg, about 2200 mg, about 2225 mg,about 2250 mg, about 2275 mg, about 2300 mg, about 2325 mg, about 2350mg, about 2375 mg, about 2400 mg, about 2425 mg, about 2450 mg, about2475 mg or about 2500 mg.

In another embodiment, a composition useful in accordance with theinvention contains not more than about 10%, not more than about 9%, notmore than about 8%, not more than about 7%, not more than about 6%, notmore than about 5%, not more than about 4%, not more than about 3%, notmore than about 2%, not more than about 1%, or not more than about 0.5%,by weight, docosahexaenoic acid (DHA) or derivative thereof such asethyl-DHA, if any. In another embodiment, a composition of the inventioncontains substantially no DHA or ethyl-DHA. In still another embodiment,a composition useful in the present invention contains no DHA orderivative thereof such as DHA-E.

In another embodiment, EPA comprises at least 70%, at least 80%, atleast 90%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100%, by weight, of all fatty acids present in acomposition according to the invention.

In another embodiment, a composition useful in accordance with theinvention contains less than 10%, less than 9%, less than 8%, less than7%, less than 6%, less than 5%, less than 4%, less than 3%, less than2%, less than 1%, less than 0.5% or less than 0.25%, by weight of thetotal composition or by weight of the total fatty acid content, of anyfatty acid or derivative thereof other than EPA. Illustrative examplesof a “fatty acid other than EPA” include linolenic acid (LA),arachidonic acid (AA), docosahexaenoic acid (DHA), alpha-linolenic acid(ALA), stearadonic acid (STA), eicosatrienoic acid (ETA) and/ordocosapentaenoic acid (DPA). In another embodiment, a composition usefulin accordance with the invention contains about 0.1% to about 4%, about0.5% to about 3%, or about 1% to about 2%, by weight, of total fattyacids other than EPA and/or DHA.

In another embodiment, a composition in accordance with the inventionhas one or more of the following features: (a) eicosapentaenoic acidethyl ester represents at least about 96%, at least about 97%, or atleast about 98%, by weight, of all fatty acids present in thecomposition; (b) the composition contains not more than about 4%, notmore than about 3%, or not more than about 2%, by weight, of total fattyacids other than eicosapentaenoic acid ethyl ester; (c) the compositioncontains not more than about 0.6%, not more than about 0.5%, or not morethan about 0.4% of any individual fatty acid other than eicosapentaenoicacid ethyl ester; (d) the composition has a refractive index (20° C.) ofabout 1 to about 2, about 1.2 to about 1.8 or about 1.4 to about 1.5;(e) the composition has a specific gravity (20° C.) of about 0.8 toabout 1.0, about 0.85 to about 0.95 or about 0.9 to about 0.92; (e) thecomposition contains not more than about 20 ppm, not more than about 15ppm or not more than about 10 ppm heavy metals, (f) the compositioncontains not more than about 5 ppm, not more than about 4 ppm, not morethan about 3 ppm, or not more than about 2 ppm arsenic, and/or (g) thecomposition has a peroxide value of not more than about 5 meq/kg, notmore than about 4 meq/kg, not more than about 3 meq/kg, or not more thanabout 2 meq/kg.

In another embodiment, the invention provides a composition comprising,consisting essentially of, or consisting of at least 95%, 96% or 97%, byweight, ethyl eicosapentaenoate, about 0.2% to about 0.5% by weightethyl octadecatetraenoate, about 0.05% to about 0.25% by weight ethylnonaecapentaenoate, about 0.2% to about 0.45% by weight ethylarachidonate, about 0.3% to about 0.5% by weight ethyleicosatetraenoate, and about 0.05% to about 0.32% ethylheneicosapentaenoate. Optionally, the composition contains not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight, DHA or derivativethere of such as ethyl-DHA. In one embodiment the composition containssubstantially no or no amount of DHA or derivative there of such asethyl-DHA. The composition further optionally comprises one or moreantioxidants (e.g. tocopherol) or other impurities in an amount of notmore than about 0.5% or not more than 0.05%. In another embodiment, thecomposition comprises about 0.05% to about 0.4%, for example about 0.2%by weight tocopherol. In another embodiment, about 500 mg to about 1 gof the composition is provided in a capsule shell.

In another embodiment, the invention provides a composition comprising,consisting of or consisting essentially of at least 96% by weight ethyleicosapentaenoate, about 0.22% to about 0.4% by weight ethyloctadecatetraenoate, about 0.075% to about 0.20% by weight ethylnonaecapentaenoate, about 0.25% to about 0.40% by weight ethylarachidonate, about 0.3% to about 0.4% by weight ethyl eicosatetraenoateand about 0.075% to about 0.25% ethyl heneicosapentaenoate. Optionally,the composition contains not more than about 0.06%, about 0.05%, orabout 0.04%, by weight, DHA or derivative there of such as ethyl-DHA. Inone embodiment the composition contains substantially no or no amount ofDHA or derivative there of such as ethyl-DHA. The composition furtheroptionally comprises one or more antioxidants (e.g. tocopherol) or otherimpurities in an amount of not more than about 0.5% or not more than0.05%. In another embodiment, the composition comprises about 0.05% toabout 0.4%, for example about 0.2% by weight tocopherol. In anotherembodiment, the invention provides a dosage form comprising about 500 mgto about 1 g of the foregoing composition in a capsule shell.

In another embodiment, the invention provides a composition comprising,consisting of, or consisting essentially of at least 96%, 97% or 98%, byweight, ethyl eicosapentaenoate, about 0.25% to about 0.38% by weightethyl octadecatetraenoate, about 0.10% to about 0.15% by weight ethylnonaecapentaenoate, about 0.25% to about 0.35% by weight ethylarachidonate, about 0.31% to about 0.38% by weight ethyleicosatetraenoate, and about 0.08% to about 0.20% ethylheneicosapentaenoate. Optionally, the composition contains not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight, DHA or derivativethere of such as ethyl-DHA. In one embodiment the composition containssubstantially no or no amount of DHA or derivative there of such asethyl-DHA. The composition further optionally comprises one or moreantioxidants (e.g. tocopherol) or other impurities in an amount of notmore than about 0.5% or not more than 0.05%. In another embodiment, thecomposition comprises about 0.05% to about 0.4%, for example about 0.2%by weight tocopherol. In another embodiment, the invention provides adosage form comprising about 500 mg to about 1 g of the foregoingcomposition in a capsule shell.

In another embodiment, the invention provides a method of increasingserum, plasma and/or red blood cell (RBC) EPA levels comprisingadministering a composition as described herein to a subject in need ofsuch treatment. In one embodiment, upon orally administering acomposition as set forth herein to a subject for a period of at leastabout 5, about 10, about 15, about 20, about 25, about 30, about 35,about 40, about 42, about 45 or about 50 days, the subject exhibits atleast about a 2-fold, at least about a 3-fold, at least about a3.5-fold, at least about a 3.75-fold or at least about a 4-fold change(final absolute EPA level divided by baseline EPA level) in serum,plasma and/or RBC EPA. In one embodiment, the method comprises a step ofidentifying a patient in need of an increase in serum, plasma and/or redblood cell (RBC) EPA prior to said administration step. In a relatedembodiment, the subject has a baseline EPA plasma, serum and/or RBClevel not greater than about 50 μg/g. In another embodiment, the subjectis provided with about 2 g to about 4 g per day of said composition. Inanother embodiment, upon administering the composition to the subject asper above, the subject exhibits a decrease in DHA, AA and/or DGLAplasma, serum and/or RBC levels. In another embodiment, uponadministering the composition to the subject as per above, the subjectexhibits an increase in DPA plasma, serum and/or RBC levels. In stillanother embodiment, upon administering the composition to the subject asper above, DHA plasma, serum and/or RBC levels decrease by at least 16%,DGLA plasma, serum and/or RBC levels decrease by at least 31%, AAplasma, serum and/or RBC levels decrease by at least 20%, and/or DPAplasma, serum and/or RBC levels increase by greater than 130%.

In another embodiment, the invention provides a method of increasingserum, plasma and/or red blood cell (RBC) EPA levels comprisingadministering a composition as described herein to a subject in need ofincreased serum, plasma and/or RBC EPA levels. In a related embodiment,upon administering the composition to the subject for a period of atleast about 5, about 10, about 15, about 20, about 25, about 30, about35, about 40, about 42, about 45, or about 50 days, the subject exhibitsat least about a 100%, at least about a 150%, at least about a 200%, atleast about a 250%, at least about a 300%, at least about a 350% or atleast about a 400% increase (change in EPA level from baseline dividedby baseline EPA level) in plasma, serum and/or RBC EPA levels comparedto baseline or placebo control. In a related embodiment, the subject hasa baseline EPA plasma, serum and/or RBC level not greater than about 50μg/g. In another embodiment, the subject is provided with about 2 g toabout 4 g per day of said composition. In another embodiment, uponadministering the composition to the subject as per above, the subjectexhibits a decrease in DHA, AA and/or DGLA plasma, serum and/or RBClevels. In another embodiment, upon administering the composition to thesubject as per above, the subject exhibits an increase in DPA plasma,serum and/or RBC levels. In still another embodiment, upon administeringthe composition to the subject as per above, DHA plasma, serum and/orRBC levels decrease by at least 16%, DGLA plasma, serum and/or RBClevels decrease by at least 31%, AA plasma, serum and/or RBC levelsdecrease by at least 20%, and/or DPA plasma, serum and/or RBC levelsincrease by greater than 130%.

In a related embodiment, upon orally administering about 2 to about 4 gper day of a composition as set forth herein to a subject for a periodof at least about 5, about 10, about 15, about 20, about 25, about 30,about 35, about 40, about 45 or about 50 days, the subject exhibits atleast about a 10 μg/g increase, at least about a 15 μg/g increase, atleast about a 20 μg/g increase, at least about a 25 μg/g increase, atleast about a 30 μg/g increase, at least about a 35 μg/g increase, atleast about a 40 μg/g increase, at least about a 45 μg/g increase, atleast about a 50 μg/g increase, at least about a 75 μg/g increase, atleast about a 100 μg/g increase, or at least about a 150 μg/g increasein serum, plasma and/or RBC EPA compared to baseline or placebo control.In another embodiment, upon administering the composition to the subjectas per above, the subject exhibits a decrease in DHA, AA and/or DGLAplasma, serum and/or RBC levels. In another embodiment, uponadministering the composition to the subject as per above, the subjectexhibits an increase in DPA plasma, serum and/or RBC levels. In stillanother embodiment, upon administering the composition to the subject asper above, DHA plasma, serum and/or RBC levels decrease by at least 16%,DGLA plasma, serum and/or RBC levels decrease by at least 31%, AAplasma, serum and/or RBC levels decrease by at least 20%, and/or DPAplasma, serum and/or RBC levels increase by greater than 130%.

In another embodiment, the subject has not been on an omega-3 fatty acidtherapy or supplement for at least 2 weeks, 3 weeks, 4 weeks, 6 weeks or12 weeks prior to initiating therapy as described herein.

In one embodiment, the invention provides a composition for use in thetreatment of, or a method for treatment and/or prevention ofcardiovascular-related diseases comprising administering to a subject inneed of such treatment or prevention a composition as set forth herein.The term “cardiovascular-related disease” herein refers to any diseaseor disorder of the heart or blood vessels (i.e. arteries and veins) orany symptom thereof. Non-limiting examples of cardiovascular-relateddisease and disorders include hypertriglyceridemia,hypercholesterolemia, mixed dyslipidemia, coronary heart disease,vascular disease, stroke, atherosclerosis, arrhythmia, hypertension,myocardial infarction, and other cardiovascular events.

The term “treatment” in relation a given disease or disorder, includes,but is not limited to, inhibiting the disease or disorder, for example,arresting the development of the disease or disorder; relieving thedisease or disorder, for example, causing regression of the disease ordisorder; or relieving a condition caused by or resulting from thedisease or disorder, for example, relieving, preventing or treatingsymptoms of the disease or disorder. The term “prevention” in relationto a given disease or disorder means: preventing the onset of diseasedevelopment if none had occurred, preventing the disease or disorderfrom occurring in a subject that may be predisposed to the disorder ordisease but has not yet been diagnosed as having the disorder ordisease, and/or preventing further disease/disorder development ifalready present.

In one embodiment, the present invention provides a method of bloodlipid therapy comprising administering to a subject or subject group inneed thereof a pharmaceutical composition as described herein. Inanother embodiment, the subject or subject group hashypertriglyceridemia, hypercholesterolemia, mixed dyslipidemia and/orvery high triglycerides.

In another embodiment, the subject or subject group being treated has abaseline triglyceride level (or mean or median baseline triglyceridelevel in the case of a subject group), fed or fasting, of about 200mg/dl to about 500 mg/dl. In another embodiment, the subject or subjectgroup has a baseline LDL-C level (or mean or median baseline LDL-Clevel), despite statin therapy, of about 40 mg/dl to about 100 mg/dl.

In one embodiment, the subject or subject group being treated inaccordance with methods of the invention is on concomitant statintherapy, for example atorvastatin, rosuvastatin or simvastatin therapy(with or without ezetimibe). In another embodiment, the subject is onconcomitant stable statin therapy at time of initiation of ultra-pureEPA therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention has a body mass index (BMI ormean BMI) of not more than about 45 kg/m².

In one embodiment, the invention provides a method of loweringtriglycerides in a subject on stable statin therapy having baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl, the methodcomprising administering to the subject a pharmaceutical compositioncomprising about 1 g to about 4 g of EPA (e.g. ultra-pure EPA), whereinupon administering the composition to the subject daily for a period ofabout 12 weeks the subject exhibits at least 10%, at least 15%, at least20%, at least 25%, at least 30%, at least 35%, at least 40%, at least45%, at least 50%, at least 55%, at least 60%, at least 65%, at least70%, or at least 75% lower fasting triglycerides than a control subjectmaintained on stable statin therapy without concomitant ultra-pure EPAfor a period of about 12 weeks, wherein the control subject also hasbaseline fasting triglycerides of about 200 mg/dl to about 500 mg/dl.The term “stable statin therapy” herein means that the subject, subjectgroup, control subject or control subject group in question has beentaking a stable daily dose of a statin (e.g. atorvastatin, rosuvastatinor simvastatin) for at least 4 weeks prior to the baseline fastingtriglyceride measurement (the “qualifying period”). For example, asubject or control subject on stable statin therapy would receive aconstant daily (i.e. the same dose each day) statin dose for at least 4weeks immediately prior to baseline fasting triglyceride measurement. Inone embodiment, the subject's and control subject's LDL-C is maintainedbetween about 40 mg/dl and about 100 mg/dl during the qualifying period.The subject and control subject are then continued on their stablestatin dose for the 12 week period post baseline.

In one embodiment, the statin is administered to the subject and thecontrol subject in an amount of about 1 mg to about 500 mg, about 5 mgto about 200 mg, or about 10 mg to about 100 mg, for example about 1 mg,about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg,about 8 mg, about 9 mg, or about 10 mg; about 15 mg, about 20 mg, about25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg,about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 175mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425mg, about 450 mg, about 475 mg, or about 500 mg. In another embodiment,the subject (and optionally the control subject) has a baseline LDL-Clevel, despite stable statin therapy, of about 40 mg/dl to about 100mg/dl. In another embodiment, the subject and/or control subject has abody mass index (BMI; or mean BMI) of not more than about 45 kg/m².

In another embodiment, the invention provides a method of loweringtriglycerides in a subject group on stable statin therapy having meanbaseline fasting triglycerides of about 200 mg/dl to about 500 mg/dl,the method comprising administering to members of the subject group apharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA per day, wherein upon administering the composition tothe members of the subject group daily for a period of about 12 weeksthe subject group exhibits at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75% lower mean fasting triglycerides than a control subject groupmaintained on stable statin therapy without concomitant ultra-pure EPAfor a period of about 12 weeks, wherein the control subject group alsohas mean baseline fasting triglycerides of about 200 mg/dl to about 500mg/dl. In a related embodiment, the stable statin therapy will besufficient such that the subject group has a mean LDL-C level about atleast about 40 mg/dl and not more than about 100 mg/dl for the 4 weeksimmediately prior to the baseline fasting triglyceride measurement.

In another embodiment, the invention provides a method of loweringtriglycerides in subject group on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to members of the subjectgroup a pharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to members ofthe subject group daily for a period of about 12 weeks the subject groupexhibits (a) at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 55%, at least 60%, at least 65%, at least 70%, at least 75% lowermean fasting triglycerides by comparison with a control subject groupmaintained on stable statin therapy without concomitant ultra-pure EPAfor a period of about 12 weeks, and (b) no increase in mean serum LDL-Clevels compared to baseline or placebo control, wherein the controlsubject also has mean baseline fasting triglycerides of about 200 mg/dlto about 500 mg/dl.

In another embodiment, the invention provides a method of loweringtriglycerides in subject on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to the subject apharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to thesubject daily for a period of about 12 weeks the subject exhibits (a) atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, or at least 75% lower fastingtriglycerides by comparison with a control subject maintained on stablestatin therapy without concomitant ultra-pure EPA for a period of about12 weeks and (b) no increase in serum LDL-C levels compared to baselineor placebo control, wherein the control subject also has baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl.

In another embodiment, the invention provides a method of loweringtriglycerides in subject group on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to members of the subjectgroup a pharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to themembers of the subject group daily for a period of about 12 weeks thesubject group exhibits (a) at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75% lower mean fasting triglycerides and (b) at least 5%, at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45% or at least 50% lower mean plasma orserum LDL-C levels by comparison with a control subject group maintainedon stable statin therapy without concomitant ultra-pure EPA for a periodof about 12 weeks, wherein the control subject also has mean baselinefasting triglycerides of about 200 mg/dl to about 500 mg/dl.

In another embodiment, the invention provides a method of loweringtriglycerides in subject group on stable statin therapy and having meanbaseline fasting triglyceride level of about 200 mg/dl to about 500mg/dl, the method comprising administering to members of the subjectgroup a pharmaceutical composition comprising about 1 g to about 4 g ofultra-pure EPA, wherein upon administering the composition to themembers of the subject group daily for a period of about 12 weeks thesubject group exhibits (a) at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75% lower mean fasting triglycerides and (b) at least 5%, at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45% or at least 50% lower mean plasma orserum LDL-C levels by comparison with a control subject group maintainedon stable statin therapy without concomitant ultra-pure EPA for a periodof about 12 weeks, wherein the control subject group also has meanbaseline fasting triglycerides of about 200 mg/dl to about 500 mg/dl.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of free total fatty acid (or mean thereof) notgreater than about 300 nmol/ml, not greater than about 250 nmol/ml, notgreater than about 200 nmol/ml, not greater than about 150 nmol/ml, notgreater than about 100 nmol/ml, or not greater than about 50 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof in the case of asubject group) not greater than about 0.70 nmol/ml, not greater thanabout 0.65 nmol/ml, not greater than about 0.60 nmol/ml, not greaterthan about 0.55 nmol/ml, not greater than about 0.50 nmol/ml, notgreater than about 0.45 nmol/ml, or not greater than about 0.40 nmol/ml.In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a baseline fastingplasma level (or mean thereof) of free EPA, expressed as a percentage oftotal free fatty acid, of not more than about 3%, not more than about2.5%, not more than about 2%, not more than about 1.5%, not more thanabout 1%, not more than about 0.75%, not more than about 0.5%, not morethan about 0.25%, not more than about 0.2% or not more than about 0.15%.In one such embodiment, free plasma EPA and/or total fatty acid levelsare determined prior to initiating therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof) not greater thanabout 1 nmol/ml, not greater than about 0.75 nmol/ml, not greater thanabout 0.50 nmol/ml, not greater than about 0.4 nmol/ml, not greater thanabout 0.35 nmol/ml, or not greater than about 0.30 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineplasma, serum or red blood cell membrane EPA level not greater thanabout 150 μg/ml, not greater than about 125 μg/ml, not greater thanabout 100 μg/ml, not greater than about 95 μg/ml, not greater than about75 μg/ml, not greater than about 60 μg/ml, not greater than about 50μg/ml, not greater than about 40 μg/ml, not greater than about 30 μg/ml,or not greater than about 25 μg/ml.

In another embodiment, methods of the present invention comprise a stepof measuring the subject's (or subject group's mean) baseline lipidprofile prior to initiating therapy. In another embodiment, methods ofthe invention comprise the step of identifying a subject or subjectgroup having one or more of the following: baseline non-HDL-C value ofabout 200 mg/dl to about 400 mg/dl, for example at least about 210mg/dl, at least about 220 mg/dl, at least about 230 mg/dl, at leastabout 240 mg/dl, at least about 250 mg/dl, at least about 260 mg/dl, atleast about 270 mg/dl, at least about 280 mg/dl, at least about 290mg/dl, or at least about 300 mg/dl; baseline total cholesterol value ofabout 250 mg/dl to about 400 mg/dl, for example at least about 260mg/dl, at least about 270 mg/dl, at least about 280 mg/dl or at leastabout 290 mg/dl; baseline vLDL-C value of about 140 mg/dl to about 200mg/dl, for example at least about 150 mg/dl, at least about 160 mg/dl,at least about 170 mg/dl, at least about 180 mg/dl or at least about 190mg/dl; baseline HDL-C value of about 10 to about 100 mg/dl, for examplenot more than about 90 mg/dl not, not more than about 80 mg/dl, not morethan about 70 mg/dl, not more than about 60 mg/dl, not more than about60 mg/dl, not more than about 50 mg/dl, not more than about 40 mg/dl,not more than about 35 mg/dl, not more than about 30 mg/dl, not morethan about 25 mg/dl, not more than about 20 mg/dl, or not more thanabout 15 mg/dl; and/or baseline LDL-C value of about 30 to about 300mg/dl, for example not less than about 40 mg/dl, not less than about 50mg/dl, not less than about 60 mg/dl, not less than about 70 mg/dl, notless than about 90 mg/dl or not less than about 90 mg/dl.

In a related embodiment, upon treatment in accordance with the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits one or more of the following outcomes:

(a) reduced triglyceride levels compared to baseline or placebo control;

(b) reduced Apo B levels compared to baseline or placebo control;

(c) increased HDL-C levels compared to baseline or placebo control;

(d) no increase in LDL-C levels compared to baseline or placebo control;

(e) a reduction in LDL-C levels compared to baseline or placebo control;

(f) a reduction in non-HDL-C levels compared to baseline or placebocontrol;

(g) a reduction in vLDL levels compared to baseline or placebo control;

(h) an increase in apo A-I levels compared to baseline or placebocontrol;

(i) an increase in apo A-I/apo B ratio compared to baseline or placebocontrol;

(j) a reduction in lipoprotein a levels compared to baseline or placebocontrol;

(k) a reduction in LDL particle number compared to baseline or placebocontrol;

(l) a reduction in LDL size compared to baseline or placebo control;

(m) a reduction in remnant-like particle cholesterol compared tobaseline or placebo control;

(n) a reduction in oxidized LDL compared to baseline or placebo control;

(o) a reduction in fasting plasma glucose (FPG) compared to baseline orplacebo control;

(p) a reduction in hemoglobin A_(1c) (HbA_(1c)) compared to baseline orplacebo control;

(q) a reduction in homeostasis model insulin resistance compared tobaseline or placebo control;

(r) a reduction in lipoprotein associated phospholipase A2 compared tobaseline or placebo control;

(s) a reduction in intracellular adhesion molecule-1 compared tobaseline or placebo control;

(t) a reduction in interleukin-2 compared to baseline or placebocontrol;

(u) a reduction in plasminogen activator inhibitor-1 compared tobaseline or placebo control;

(v) a reduction in high sensitivity C-reactive protein (hsCRP) comparedto baseline or placebo control;

(w) an increase in plasma or serum phospholipid EPA compared to baselineor placebo control;

(x) an increase in red blood cell membrane EPA compared to baseline orplacebo control; and/or

(y) a reduction or increase in one or more of plasma, serum phospholipidand/or red blood cell content of docosahexaenoic acid (DHA),docosapentaenoic acid (DPA), arachidonic acid (AA), palmitic acid (PA),staeridonic acid (SA) or oleic acid (OA) compared to baseline or placebocontrol.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(y) above priorto dosing the subject or subject group. In another embodiment, themethods comprise administering a composition as disclosed herein to thesubject after baseline levels of one or more markers set forth in(a)-(y) are determined, and subsequently taking an additionalmeasurement of said one or more markers.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more, any 24 or more, or all 25 of outcomes (a)-(y) describedimmediately above.

In another embodiment, upon treatment with a composition of the presentinvention, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in triglyceride level of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or placebocontrol;

(b) a less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in non-HDL-C levels or areduction in non-HDL-C levels of at least about 1%, at least about 3%,at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55% or at least about 75% (actual % change or median % change) ascompared to baseline or placebo control;

(c) an increase in HDL-C levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or placebocontrol;

(d) a less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in LDL-C levels or areduction in LDL-C levels of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 55% or at leastabout 75% (actual % change or median % change) as compared to baselineor placebo control;

(e) a decrease in Apo B levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55% or at least about 75% (actual %change or median % change) as compared to baseline or placebo control;

(f) a reduction in vLDL levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, or at least about 100% (actual % change or median %change) compared to baseline or placebo control;

(g) an increase in apo A-I levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or placebo control;

(h) an increase in apo A-I/apo B ratio of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or placebo control;

(i) a reduction in lipoprotein(a) levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or placebo control;

(j) a reduction in mean LDL particle number of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or placebo control;

(k) an increase in mean LDL particle size of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or placebo control;

(l) a reduction in remnant-like particle cholesterol of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or placebo control;

(m) a reduction in oxidized LDL of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or placebo control;

(n) a reduction in fasting plasma glucose (FPG) of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or placebo control;

(o) a reduction in hemoglobin A_(1c) (HbA_(1c)) of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, or at least about 50% (actual % change or median %change) compared to baseline or placebo control;

(p) a reduction in homeostasis model index insulin resistance of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline orplacebo control;

(q) a reduction in lipoprotein associated phospholipase A2 of at leastabout 5%, at least about 10%, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, or at least about 100%(actual % change or median % change) compared to baseline or placebocontrol;

(r) a reduction in intracellular adhesion molecule-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or placebo control;

(s) a reduction in interleukin-2 of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or placebo control;

(t) a reduction in plasminogen activator inhibitor-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or placebo control;

(u) a reduction in high sensitivity C-reactive protein (hsCRP) of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline orplacebo control;

(v) an increase in plasma, serum phospholipids or RBC EPA of at leastabout 5%, at least about 10%, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, at least about 100%, atleast about 200% or at least about 400% (actual % change or median %change) compared to baseline or placebo control;

(w) an increase in plasma, serum phospholipid and/or RBC membrane EPA ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 100%, at least about 200%, or at least about 400% (actual % changeor median % change) compared to baseline or placebo control;

(x) a reduction or increase in one or more of plasma, serum phospholipidand/or RBC DHA, DPA, AA, PA and/or OA of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or placebocontrol; and/or

(y) a reduction in total cholesterol of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or placebocontrol.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(y) prior todosing the subject or subject group. In another embodiment, the methodscomprise administering a composition as disclosed herein to the subjectafter baseline levels of one or more markers set forth in (a)-(y) aredetermined, and subsequently taking a second measurement of the one ormore markers as measured at baseline for comparison thereto.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more of, any 24 or more of, or all 26 or more of outcomes (a)-(y)described immediately above.

Parameters (a)-(y) can be measured in accordance with any clinicallyacceptable methodology. For example, triglycerides, total cholesterol,HDL-C and fasting blood sugar can be sample from serum and analyzedusing standard photometry techniques. VLDL-TG, LDL-C and VLDL-C can becalculated or determined using serum lipoprotein fractionation bypreparative ultracentrifugation and subsequent quantitative analysis byrefractometry or by analytic ultracentrifugal methodology. Apo A1, Apo Band hsCRP can be determined from serum using standard nephelometrytechniques. Lipoprotein (a) can be determined from serum using standardturbidimetric immunoassay techniques. LDL particle number and particlesize can be determined using nuclear magnetic resonance (NMR)spectrometry. Remnants lipoproteins and LDL-phospholipase A2 can bedetermined from EDTA plasma or serum and serum, respectively, usingenzymatic immunoseparation techniques. Oxidized LDL, intercellularadhesion molecule-1 and interleukin-2 levels can be determined fromserum using standard enzyme immunoassay techniques. These techniques aredescribed in detail in standard textbooks, for example TietzFundamentals of Clinical Chemistry, 6^(th) Ed. (Burtis, Ashwood andBorter Eds.), WB Saunders Company.

In one embodiment, subjects fast for up to 12 hours prior to bloodsample collection, for example about 10 hours.

In another embodiment, the present invention provides a method oftreating or preventing primary hypercholesterolemia and/or mixeddyslipidemia (Fredrickson Types IIa and IIb) in a patient in needthereof, comprising administering to the patient one or morecompositions as disclosed herein. In a related embodiment, the presentinvention provides a method of reducing triglyceride levels in a subjector subjects when treatment with a statin or niacin extended-releasemonotherapy is considered inadequate (Frederickson type IVhyperlipidemia).

In another embodiment, the present invention provides a method oftreating or preventing risk of recurrent nonfatal myocardial infarctionin a patient with a history of myocardial infarction, comprisingadministering to the patient one or more compositions as disclosedherein.

In another embodiment, the present invention provides a method ofslowing progression of or promoting regression of atheroscleroticdisease in a patient in need thereof, comprising administering to asubject in need thereof one or more compositions as disclosed herein.

In another embodiment, the present invention provides a method oftreating or preventing very high serum triglyceride levels (e.g. TypesIV and V hyperlipidemia) in a patient in need thereof, comprisingadministering to the patient one or more compositions as disclosedherein.

In one embodiment, a composition of the invention is administered to asubject in an amount sufficient to provide a daily dose of ethyleicosapentaenoic acid of about 1 mg to about 10,000 mg, 25 about 5000mg, about 50 to about 3000 mg, about 75 mg to about 2500 mg, or about100 mg to about 1000 mg, for example about 75 mg, about 100 mg, about125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg,about 1025 mg, about 1050 mg, about 1075 mg, about 1200 mg, about 1225mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg,about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575mg, about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about1700 mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg,about 1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925mg, about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about2050 mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg,about 2175 mg, about 2200 mg, about 2225 mg, about 2250 mg, about 2275mg, about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about2400 mg, about 2425 mg, about 2450 mg, about 2475 mg or about 2500 mg.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume a traditional Westerndiet. In one embodiment, the methods of the invention include a step ofidentifying a subject as a Western diet consumer or prudent dietconsumer and then treating the subject if the subject is deemed aWestern diet consumer. The term “Western diet” herein refers generallyto a typical diet consisting of, by percentage of total calories, about45% to about 50% carbohydrate, about 35% to about 40% fat, and about 10%to about 15% protein. A Western diet may alternately or additionally becharacterized by relatively high intakes of red and processed meats,sweets, refined grains, and desserts, for example more than 50%, morethan 60% or more or 70% of total calories come from these sources.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume less than (actual oraverage) about 150 g, less than about 125 g, less than about 100 g, lessthan about 75 g, less than about 50 g, less than about 45 g, less thanabout 40 g, less than about 35 g, less than about 30 g, less than about25 g, less than about 20 g or less than about 15 g of fish per day.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume less than (actual oraverage) about 10 g, less than about 9 g, less than about 8 g, less thanabout 7 g, less than about 6 g, less than about 5 g, less than about 4g, less than about 3 g, less than about 2 g per day of omega-3 fattyacids from dietary sources.

In another embodiment, any of the methods disclosed herein are used intreatment of a subject or subjects that consume less than (actual oraverage) about 2.5 g, less than about 2 g, less than about 1.5 g, lessthan about 1 g, less than about 0.5 g, less than about 0.25 g, or lessthan about 0.2 g per day of EPA and DHA (combined) from dietary sources.

Seafood/fish allergies often manifest as hives, itching, and eczema(rash). These reactions among those with fish allergies prevent patientsfrom benefiting from the cardiovascular, anti-inflammatory,neurological, endocrine and other benefits of pharmaceutical grade fishoil derivatives by decreasing compliance. In various embodiments, theinvention provides compositions and methods as described herein whereinthe subject does not exhibit skin rash, or exhibits reduced skin rashincidence compared to placebo.

Another shortcoming of fish oil derived therapies and medicines are theincidence of eructation, or “fishy burp”. In various embodiments, theinvention provides compositions and methods as described herein whereinthe subject does not experience eructation or experiences reducedeructation compared to Lovaza therapy.

In one embodiment, a composition as described herein is administered toa subject once or twice per day. In another embodiment, 1, 2, 3 or 4capsules, each containing about 500 mg to about 1 g of a composition asdescribed herein, are administered to a subject daily. In anotherembodiment, 1 or 2 capsules, each containing about 1 g of a compositionas described herein, are administered to the subject in the morning, forexample between about 5 am and about 11 am, and 1 or 2 capsules, eachcontaining about 1 g of a composition as described herein, areadministered to the subject in the evening, for example between about 5pm and about 11 pm.

In one embodiment, a subject being treated in accordance with methods ofthe invention is not on fibrate or nitrate therapy.

In another embodiment, compositions useful in accordance with methods ofthe invention are orally deliverable. The terms “orally deliverable” or“oral administration” herein include any form of delivery of atherapeutic agent or a composition thereof to a subject wherein theagent or composition is placed in the mouth of the subject, whether ornot the agent or composition is swallowed. Thus “oral administration”includes buccal and sublingual as well as esophageal administration. Inone embodiment, the composition is present in a capsule, for example asoft gelatin capsule.

A composition for use in accordance with the treatments of invention canbe formulated as one or more dosage units. The terms “dose unit” and“dosage unit” herein refer to a portion of a pharmaceutical compositionthat contains an amount of a therapeutic agent suitable for a singleadministration to provide a therapeutic effect. Such dosage units may beadministered one to a plurality (i.e. 1 to about 10, 1 to 8, 1 to 6, 1to 4 or 1 to 2) of times per day, or as many times as needed to elicit atherapeutic response.

In another embodiment, the invention provides use of any compositiondescribed herein for treating moderate to severe hypertriglyceridemia ina subject in need thereof, comprising: providing a subject having afasting baseline triglyceride level of about 500 mg/dl to about 1500mg/dl and administering to the subject a pharmaceutical composition asdescribed herein. In one embodiment, the composition comprises about 1 gto about 4 g of eicosapentaenoic acid ethyl ester, wherein thecomposition contains substantially no docosahexaenoic acid.

In one embodiment, the present invention provides a method of bloodlipid therapy comprising administering to a subject or subject group inneed thereof a pharmaceutical composition as described herein. Inanother embodiment, the subject or subject group hashypertriglyceridemia, hypercholesterolemia, mixed dyslipidemia and/orvery high triglycerides.

In another embodiment, the subject or subject group being treated has abaseline triglyceride level (or median baseline triglyceride level inthe case of a subject group), fed or fasting, of at least about 300mg/dl, at least about 400 mg/dl, at least about 500 mg/dl, at leastabout 600 mg/dl, at least about 700 mg/dl, at least about 800 mg/dl, atleast about 900 mg/dl, at least about 1000 mg/dl, at least about 1100mg/dl, at least about 1200 mg/dl, at least about 1300 mg/dl, at leastabout 1400 mg/dl, or at least about 1500 mg/dl, for example about 400mg/dl to about 2500 mg/dl, about 450 mg/dl to about 2000 mg/dl or about500 mg/dl to about 1500 mg/dl.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention has previously been treatedwith Lovaza® and has experienced an increase in, or no decrease in,LDL-C levels and/or non-HDL-C levels. In one such embodiment, Lovaza®therapy is discontinued and replaced by a method of the presentinvention.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof in the case of asubject group) not greater than about 0.70 nmol/ml, not greater thanabout 0.65 nmol/ml, not greater than about 0.60 nmol/ml, not greaterthan about 0.55 nmol/ml, not greater than about 0.50 nmol/ml, notgreater than about 0.45 nmol/ml, or not greater than about 0.40 nmol/ml.In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a baseline fastingplasma level (or mean thereof) of free EPA, expressed as a percentage oftotal free fatty acid, of not more than about 3%, not more than about2.5%, not more than about 2%, not more than about 1.5%, not more thanabout 1%, not more than about 0.75%, not more than about 0.5%, not morethan about 0.25%, not more than about 0.2% or not more than about 0.15%.In one such embodiment, free plasma EPA and/or total fatty acid levelsare determined prior to initiating therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of total fatty acid (or mean thereof) not greaterthan about 250 nmol/ml, not greater than about 200 nmol/ml, not greaterthan about 150 nmol/ml, not greater than about 100 nmol/ml, or notgreater than about 50 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineplasma, serum or red blood cell membrane EPA level not greater thanabout 70 μg/ml, not greater than about 60 μg/ml, not greater than about50 μg/ml, not greater than about 40 μg/ml, not greater than about 30μg/ml, or not greater than about 25 μg/ml.

In another embodiment, methods of the present invention comprise a stepof measuring the subject's (or subject group's mean) baseline lipidprofile prior to initiating therapy. In another embodiment, methods ofthe invention comprise the step of identifying a subject or subjectgroup having one or more of the following: baseline non-HDL-C value ofabout 200 mg/dl to about 400 mg/dl, for example at least about 210mg/dl, at least about 220 mg/dl, at least about 230 mg/dl, at leastabout 240 mg/dl, at least about 250 mg/dl, at least about 260 mg/dl, atleast about 270 mg/dl, at least about 280 mg/dl, at least about 290mg/dl, or at least about 300 mg/dl; baseline total cholesterol value ofabout 250 mg/dl to about 400 mg/dl, for example at least about 260mg/dl, at least about 270 mg/dl, at least about 280 mg/dl or at leastabout 290 mg/dl; baseline vLDL-C value of about 140 mg/dl to about 200mg/dl, for example at least about 150 mg/dl, at least about 160 mg/dl,at least about 170 mg/dl, at least about 180 mg/dl or at least about 190mg/dl; baseline HDL-C value of about 10 to about 60 mg/dl, for examplenot more than about 40 mg/dl, not more than about 35 mg/dl, not morethan about 30 mg/dl, not more than about 25 mg/dl, not more than about20 mg/dl, or not more than about 15 mg/dl; and/or baseline LDL-C valueof about 50 to about 300 mg/dl, for example not less than about 100mg/dl, not less than about 90 mg/dl, not less than about 80 mg/dl, notless than about 70 mg/dl, not less than about 60 mg/dl or not less thanabout 50 mg/dl.

In one embodiment, compositions of the invention are packaged in blisterpacks. In another embodiment, the blister packs comprise PCTFE (forexample 50μ) laminated with water based adhesive to clear PVC (forexample 190μ) which are heat sealed to aluminum foil).

In a related embodiment, upon treatment in accordance with the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits one or more of the following outcomes:

(a) reduced triglyceride levels compared to baseline or a placebo arm;

(b) reduced Apo B levels compared to baseline or a placebo arm;

(c) increased HDL-C levels compared to baseline or a placebo arm;

(d) no increase in LDL-C levels compared to baseline or a placebo arm;

(e) a reduction in LDL-C levels compared to baseline or a placebo arm;

(f) a reduction in non-HDL-C levels compared to baseline or a placeboarm;

(g) a reduction in vLDL levels compared to baseline or a placebo arm;

(h) an increase in apo A-I levels compared to baseline or a placebo arm;

(i) an increase in apo A-I/apo B ratio compared to baseline or a placeboarm;

(j) a reduction in lipoprotein A levels compared to baseline or aplacebo arm;

(k) a reduction in LDL particle number compared to baseline or a placeboarm;

(l) an increase in mean LDL size compared to baseline or a placebo arm;

(m) a reduction in remnant-like particle cholesterol compared tobaseline or a placebo arm;

(n) a reduction in oxidized LDL compared to baseline or a placebo arm;

(o) no change or a reduction in fasting plasma glucose (FPG) compared tobaseline or a placebo arm;

(p) a reduction in hemoglobin A_(1c) (HbA_(1c)) compared to baseline ora placebo arm;

(q) a reduction in homeostasis model insulin resistance compared tobaseline or a placebo arm;

(r) a reduction in lipoprotein associated phospholipase A2 compared tobaseline or a placebo arm;

(s) a reduction in intracellular adhesion molecule-1 compared tobaseline or a placebo arm;

(t) a reduction in interleukin-6 compared to baseline or a placebo arm;

(u) a reduction in plasminogen activator inhibitor-1 compared tobaseline or a placebo arm;

(v) a reduction in high sensitivity C-reactive protein (hsCRP) comparedto baseline or a placebo arm;

(w) an increase in serum phospholipid EPA compared to baseline or aplacebo arm;

(x) an increase in red blood cell membrane EPA compared to baseline or aplacebo arm; and/or

(y) a reduction or increase in one or more of serum phospholipid and/orred blood cell content of docosahexaenoic acid (DHA), docosapentaenoicacid (DPA), arachidonic acid (AA), palmitic acid (PA), staeridonic acid(SA) or oleic acid (OA) compared to baseline or a placebo arm.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(y) above priorto dosing the subject or subject group. In another embodiment, themethods comprise administering a composition as disclosed herein to thesubject after baseline levels of one or more markers set forth in(a)-(y) are determined, and subsequently taking an additionalmeasurement of said one or more markers.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more, any 24 or more, or all 25 of outcomes (a)-(y) describedimmediately above.

In another embodiment, upon treatment with a composition of the presentinvention, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in triglyceride level of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or aplacebo arm;

(b) a less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in non-HDL-C levels or areduction in non-HDL-C levels of at least about 1%, at least about 3%,at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55% or at least about 75% (actual % change or median % change) ascompared to baseline or a placebo arm;

(c) substantially no change, no change or an increase in HDL-C levels ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55% or at least about 75% (actual % change or median % change) ascompared to baseline or a placebo arm;

(d) a less than 60% increase, less than 50% increase, less than 40%increase, less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in LDL-C levels or areduction in LDL-C levels of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 55% or at leastabout 75% (actual % change or median % change) as compared to baselineor a placebo arm;

(e) a decrease in Apo B levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55% or at least about 75% (actual %change or median % change) as compared to baseline or a placebo arm;

(f) a reduction in vLDL levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, or at least about 100% (actual % change or median %change) compared to baseline or a placebo arm;

(g) an increase in apo A-I levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or a placebo arm;

(h) an increase in apo A-I/apo B ratio of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or a placebo arm;

(i) a reduction in lipoprotein(a) levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or a placebo arm;

(j) a reduction in mean LDL particle number of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or a placebo arm;

(k) an increase in mean LDL particle size of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or a placebo arm;

(l) a reduction in remnant-like particle cholesterol of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or a placebo arm;

(m) a reduction in oxidized LDL of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or a placebo arm;

(n) substantially no change, no change or a reduction in fasting plasmaglucose (FPG) of at least about 5%, at least about 10%, at least about15%, at least about 20%, at least about 25%, at least about 30%, atleast about 35%, at least about 40%, at least about 45%, at least about50%, or at least about 100% (actual % change or median % change)compared to baseline or a placebo arm;

(o) substantially no change, no change or a reduction in hemoglobinA_(1c) (HbA_(1c)) of at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, or at leastabout 50% (actual % change or median % change) compared to baseline or aplacebo arm;

(p) a reduction in homeostasis model index insulin resistance of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline ora placebo arm;

(q) a reduction in lipoprotein associated phospholipase A2 of at leastabout 5%, at least about 10%, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, or at least about 100%(actual % change or median % change) compared to baseline or a placeboarm;

(r) a reduction in intracellular adhesion molecule-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or a placebo arm;

(s) a reduction in interleukin-6 of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or a placebo arm;

(t) a reduction in plasminogen activator inhibitor-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline;

(u) a reduction in high sensitivity C-reactive protein (hsCRP) of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline ora placebo arm;

(v) an increase in serum, plasma and/or RBC EPA of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, at least about 100%, at least about200% or at least about 400% (actual % change or median % change)compared to baseline or a placebo arm;

(w) an increase in serum phospholipid and/or red blood cell membrane EPAof at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 100%, at least about 200%, or at least about 400% (actual % changeor median % change) compared to baseline or a placebo arm;

(x) a reduction or increase in one or more of serum phospholipid and/orred blood cell DHA, DPA, AA, PA and/or OA of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or a placeboarm; and/or

(y) a reduction in total cholesterol of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or a placeboarm.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(y) prior todosing the subject or subject group. In another embodiment, the methodscomprise administering a composition as disclosed herein to the subjectafter baseline levels of one or more markers set forth in (a)-(y) aredetermined, and subsequently taking a second measurement of the one ormore markers as measured at baseline for comparison thereto.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more of, any 24 or more of, or all 25 of outcomes (a)-(y)described immediately above.

Parameters (a)-(y) can be measured in accordance with any clinicallyacceptable methodology. For example, triglycerides, total cholesterol,HDL-C and fasting blood sugar can be sample from serum and analyzedusing standard photometry techniques. VLDL-TG, LDL-C and VLDL-C can becalculated or determined using serum lipoprotein fractionation bypreparative ultracentrifugation and subsequent quantitative analysis byrefractometry or by analytic ultracentrifugal methodology. Apo A1, Apo Band hsCRP can be determined from serum using standard nephelometrytechniques. Lipoprotein (a) can be determined from serum using standardturbidimetric immunoassay techniques. LDL particle number and particlesize can be determined using nuclear magnetic resonance (NMR)spectrometry. Remnants lipoproteins and LDL-phospholipase A2 can bedetermined from EDTA plasma or serum and serum, respectively, usingenzymatic immunoseparation techniques. Oxidized LDL, intercellularadhesion molecule-1 and interleukin-2 levels can be determined fromserum using standard enzyme immunoassay techniques. These techniques aredescribed in detail in standard textbooks, for example TietzFundamentals of Clinical Chemistry, 6^(th) Ed. (Burtis, Ashwood andBorter Eds.), WB Saunders Company.

In another embodiment, the invention provides a method of treatinghypertriglyceridemia in a subject that is allergic to or hypersensitiveto fish, the method comprising identifying a subject withhypertriglyceridemia that is allergic to or hypersensitive to fish; andadministering to the subject an effective amount of a compositioncomprising eicosapentaenoic acid or a derivative thereof. In anotherembodiment, the composition contains substantially no fish allergen. Inanother embodiment, the composition contains no proteins. In anotherembodiment, the composition contains substantially no fish proteins. Inanother embodiment, the composition contains substantially no fishallergens. In another embodiment, the composition contains no fishallergens. In another embodiment, the composition contains less thanabout 5% by weight, if any, of any fatty acid other thaneicosapentaenoic acid or a derivative thereof. In another embodiment,the composition contains less than about 5% by weight of DHA or aderivative thereof. In another embodiment, the composition comprises atleast 96%, by weight, ethyl eicosapentaenoate, about 0.2% to about 0.5%by weight ethyl octadecatetraenoate, about 0.05% to about 0.25% byweight ethyl nonaceapentaenoate, about 0.2% to about 0.45% by weightethyl arachidonate, about 0.3% to about 0.5% by weight ethyleicosatetraenoate, about 0.05% to about 0.32% ethylheneicosapentaenoate, and not more than 0.05% ethyl-DHA, if any. Inanother embodiment, the composition comprises at least 96% ethyleicosapentaenoate, about 0.22% to about 0.4% by weight ethyloctadecatetraenoate, about 0.075% to about 0.20% by weight ethylnonaecapentaenoate, about 0.25% to about 0.40% by weight ethylarachidonate, about 0.3% to about 0.4% by weight ethyleicosatetraenoate, and about 0.075% to about 0.25% ethylheneicosapentaenoate. In another embodiment, the composition comprisesat least 98% ethyl eicosapentaenoate, about 0.25% to about 0.38% byweight ethyl octadecatetraenoate, about 0.10% to about 0.15% by weightethyl nonaecapentaenoate, about 0.25% to about 0.35% by weight ethylarachidonate, about 0.31% to about 0.38% by weight ethyleicosatetraenoate, and about 0.08% to about 0.20% ethylheneicosapentaenoate. In another embodiment, the composition furthercomprises tocopherol in an amount of about 0.1% to about 0.3%, byweight. In another embodiment, the eicosapentaenoic acid or derivativethereof is ethyl eicosapentaenoate. In another embodiment, the subjectis on concomitant statin therapy. In another embodiment, the subject isnot on concomitant fibrate or nitrate therapy. In another embodiment,the composition is encapsulated in a capsule.

In another embodiment, the invention provides a method of increasingserum, plasma, and/or red blood cell EPA levels in a subject that isallergic to or hypersensitive to fish, the method comprising:identifying a subject that is allergic to or hypersensitive to fish; andadministering to the subject an effective amount of a compositioncomprising eicosapentaenoic acid or a derivative thereof, wherein saidcomposition contains substantially no proteins.

In another embodiment, the invention provides a method of treating orpreventing cardiovascular-related diseases in a subject that is allergicto or hypersensitive to fish, the method comprising: identifying asubject that is allergic to or hypersensitive to fish and has or is atrisk for developing a cardiovascular-related disease; and administeringto the subject an effective amount of a composition comprisingeicosapentaenoic acid or a derivative thereof, wherein said compositioncontains substantially no proteins.

In another embodiment, the invention provides a method of blood lipidtherapy comprising: identifying a subject in need of blood lipid therapythat is allergic to or hypersensitive to fish; and administering to thesubject an effective amount of a composition comprising eicosapentaenoicacid or a derivative thereof, wherein said composition containssubstantially no proteins.

In another embodiment, the invention provides a method for reducingtriglycerides in a subject on stable statin therapy, the methodcomprising: identifying a subject that is allergic to or hypersensitiveto fish; and administering to the subject an effective amount of acomposition comprising eicosapentaenoic acid or a derivative thereof,wherein said composition contains substantially no proteins.

In another embodiment, the invention provides a method of treating orpreventing primary hypercholesterolemia and/or mixed dyslipidemia in asubject that is allergic to or hypersensitive to fish, the methodcomprising: identifying a that is allergic to or hypersensitive to fishand has or is at risk of developing primary hypercholesterolemia and/ormixed dyslipidemia; and administering to the subject an effective amountof a composition comprising eicosapentaenoic acid or a derivativethereof, wherein said composition contains substantially no proteins.

In another embodiment, the invention provides a method of reducingtriglyceride levels in a subject that is allergic to or hypersensitiveto fish after statin or niacin extended-release monotherapy is deemedinadequate, the method comprising: identifying a subject that isallergic to or hypersensitive to fish; and administering to the subjectan effective amount of a composition comprising eicosapentaenoic acid ora derivative thereof, wherein said composition contains substantially noproteins.

In another embodiment, the invention provides a method of slowingprogression of or promoting regression of atherosclerotic disease in asubject that is allergic to or hypersensitive to fish, the methodcomprising: identifying a subject with atherosclerotic disease that isallergic to or hypersensitive to fish; and administering to the subjectan effective amount of a composition comprising eicosapentaenoic acid ora derivative thereof, wherein said composition contains substantially noproteins.

In another embodiment, the invention provides a method of treating orpreventing very high serum triglyceride levels in a subject that isallergic to or hypersensitive to fish, the method comprising:identifying a subject that is allergic to or hypersensitive to fish andhas or is at risk of developing very high serum triglyceride levels; andadministering to the subject an effective amount of a compositioncomprising eicosapentaenoic acid or a derivative thereof, wherein saidcomposition contains substantially no proteins.

In another embodiment, the invention provides a method for increasingplasma and/or serum EPA levels in a subject that is allergic to orhypersensitive to fish, the method comprising: identifying a subjectthat is allergic to or hypersensitive to fish; and administering to thesubject an effective amount of a composition comprising eicosapentaenoicacid or a derivative thereof, wherein said composition containssubstantially no proteins.

In another embodiment, the invention provides a method of increasingplasma and/or serum EPA in a subject in need thereof that is allergic toor hypersensitive to fish, the method comprising: identifying a subjectthat is allergic to or hypersensitive to fish; and administering to thesubject the composition according to the present disclosure in an amountsufficient to increase plasma and/or serum EPA levels in the subject byat least about 200% compared to baseline.

In another embodiment, the invention provides a method of increasingplasma and/or serum EPA in a subject in need thereof that is allergic toor hypersensitive to fish, the method comprising: identifying a subjectthat is allergic to or hypersensitive to fish; and administering to thesubject a composition comprising at least about 96%, by weight, ofeicosapentaenoic acid or a derivative thereof in an amount sufficient toincrease plasma and/or serum EPA levels in the subject by at least about200% compared to baseline.

In another embodiment, the present invention provides a method oftreating a cardiovascular-related disease comprising administering to asubject in need thereof a composition comprising EPA and acardiovascular agent, wherein the composition contains substantially noproteins.

In another embodiment, the present invention provides a method ofreducing the risk of arterial plaque development in a subject at riskthereof, comprising: determining a risk of arterial plaque developmentin the subject by measuring an elevated level of an oxidative stressmarker in the subject; and subsequently administering to the subject apharmaceutical composition comprising EPA, DPA or a derivative thereof,wherein the pharmaceutical composition contains substantially noproteins.

In another embodiment, the present invention provides a method ofreducing the number or size of arterial plaques in a subject in needthereof comprising: determining a baseline number or size of arterialplaques in the subject; measuring an elevated level of8-hydroxy-2′-deoxyguanosine in the subject; subsequently administeringto the subject an arterial plaque-reducing amount of a pharmaceuticalcomposition comprising EPA, DPA or a derivative thereof, wherein thepharmaceutical composition contains substantially no proteins; andmeasuring a reduced number or size of arterial plaques in the subject ascompared to the baseline number or size of arterial plaques in thesubject.

In another embodiment, the present invention provides a method oftreating hypertriglyceridemia in a subject that is allergic to orhypersensitive to fish, the method comprising: (a) identifying a subjecthaving a fasting baseline triglyceride level of about 500 mg/dl to about1500 mg/dl that is allergic to or hypersensitive to fish, and (b)administering to the subject a pharmaceutical composition comprising atleast 95% all-cis eicosa-5,8,11,14,17-pentaenoic acid ethyl ester,wherein the composition is substantially free of docosahexaenoic acidethyl ester. In another embodiment, the pharmaceutical composition issubstantially free of fish proteins or fish allergens.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising eicosapentaenoic acid, or a derivative thereof,wherein the pharmaceutical composition contains less than about 1%, byweight, of proteins, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 0.5%, by weight, of proteins, ifany. In another embodiment, the pharmaceutical composition contains lessthan about 0.2%, by weight, of proteins, if any. In another embodiment,the pharmaceutical composition contains less than about 0.1%, by weight,of proteins, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 0.05%, by weight, of proteins, ifany. In another embodiment, the pharmaceutical composition contains lessthan about 0.005%, by weight, of proteins, if any. In anotherembodiment, the pharmaceutical composition contains less than about0.0005%, by weight, of proteins, if any. In another embodiment, thepharmaceutical composition contains no protein. In another embodiment,the pharmaceutical composition contains less than about 1%, by weight,of a fish allergen, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 0.5%, by weight, of a fishallergen, if any. In another embodiment, the pharmaceutical compositioncontains less than about 0.05%, by weight, of a fish allergen, if any.In another embodiment, the pharmaceutical composition contains less thanabout 0.005%, by weight, of a fish allergen, if any. In anotherembodiment, the pharmaceutical composition contains less than about0.0005%, by weight, of a fish allergen, if any. In another embodiment,the pharmaceutical composition contains no fish allergen. In anotherembodiment, the pharmaceutical composition contains less than about 5%,by weight, of any fatty acid other than eicosapentaenoic acid or aderivative thereof, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 4%, by weight, of any fatty acidother than eicosapentaenoic acid or a derivative thereof, if any. Inanother embodiment, the pharmaceutical composition contains less thanabout 3%, by weight, of any fatty acid other than eicosapentaenoic acidor a derivative thereof, if any. In another embodiment, thepharmaceutical composition contains less than about 2%, by weight, ofany fatty acid other than eicosapentaenoic acid or a derivative thereof,if any. In another embodiment, the pharmaceutical composition containsless than about 1%, by weight, of any fatty acid other thaneicosapentaenoic acid or a derivative thereof, if any. In anotherembodiment, the pharmaceutical composition contains less than about0.5%, by weight, of any fatty acid other than eicosapentaenoic acid or aderivative thereof, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 0.05%, by weight, of any fatty acidother than eicosapentaenoic acid or a derivative thereof, if any. Inanother embodiment, the pharmaceutical composition contains less thanabout 0.005%, by weight, of any fatty acid other than eicosapentaenoicacid or a derivative thereof, if any. In another embodiment, thepharmaceutical composition contains less than about 0.0005%, by weight,of any fatty acid other than eicosapentaenoic acid or a derivativethereof, if any. In another embodiment, the pharmaceutical compositioncontains less than about 0.1%, by weight, of ethyl-DHA, if any. Inanother embodiment, the pharmaceutical composition contains less thanabout 0.075%, by weight, of ethyl-DHA, if any. In another embodiment,the pharmaceutical composition contains less than about 0.05%, byweight, of ethyl-DHA, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 0.025%, by weight, of ethyl-DHA, ifany. In another embodiment, the pharmaceutical composition contains noethyl-DHA. In another embodiment, the eicosapentaenoic acid orderivative thereof is an ester of eicosapentaenoic acid. In anotherembodiment, the eicosapentaenoic acid or derivative thereof iseicosapentaenoic acid, ethyl ester.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising eicosapentaenoic acid or a derivative thereof,wherein the pharmaceutical composition contains less than about 1%, byweight, of proteins, if any, and less than about 5%, by weight, of anyfatty acid other than eicosapentaenoic acid or a derivative thereof. Inanother embodiment, the pharmaceutical composition contains less thanabout 4%, by weight, of any fatty acid other than eicosapentaenoic acidor a derivative thereof. In another embodiment, the pharmaceuticalcomposition contains less than about 3%, by weight, of any fatty acidother than eicosapentaenoic acid or a derivative thereof. In anotherembodiment, the pharmaceutical composition contains less than about 2%,by weight, of any fatty acid other than eicosapentaenoic acid or aderivative thereof. In another embodiment, the pharmaceuticalcomposition contains less than about 1%, by weight, of any fatty acidother than eicosapentaenoic acid or a derivative thereof. In anotherembodiment, the pharmaceutical composition contains less than about0.5%, by weight, of proteins, if any. In another embodiment, thepharmaceutical composition contains less than about 0.05%, by weight, ofproteins, if any. In another embodiment, the pharmaceutical compositioncontains less than about 0.005%, by weight, of proteins, if any. Inanother embodiment, the pharmaceutical composition contains less thanabout 0.0005%, by weight, of proteins, if any. In another embodiment,the pharmaceutical composition contains less than about 0.00005%, byweight, of proteins, if any. In another embodiment, the pharmaceuticalcomposition contains substantially no protein. In another embodiment,the pharmaceutical composition contains no protein. In anotherembodiment, the pharmaceutical composition contains substantially nofish protein. In another embodiment, the pharmaceutical compositioncontains no fish protein. In another embodiment, the pharmaceuticalcomposition contains substantially no fish allergen. In anotherembodiment, the pharmaceutical composition contains no fish allergen.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising eicosapentaenoic acid or a derivative thereof,octadecatetraenoic acid or a derivative thereof, nonaecapentaenoic acidor a derivative thereof, arachidonic acid or a derivative thereof,eicosatetraenoic acid or a derivative thereof, heneicosapentaenoate or aderivative thereof, and less than about 1%, by weight, of protein, ifany. In another embodiment, the pharmaceutical composition contains lessthan about 0.05%, by weight, of protein, if any. In another embodiment,the pharmaceutical composition contains less than about 0.005%, byweight, of protein, if any. In another embodiment, the pharmaceuticalcomposition contains less than about 0.0005%, by weight, of protein, ifany. In another embodiment, the pharmaceutical composition contains lessthan about 0.00005%, by weight, of protein, if any. In anotherembodiment, the pharmaceutical composition contains substantially noprotein. In another embodiment, the pharmaceutical composition containsno protein. In another embodiment, the pharmaceutical compositioncontains substantially no fish protein. In another embodiment, thepharmaceutical composition contains no fish protein. In anotherembodiment, the pharmaceutical composition contains substantially nofish allergen. In another embodiment, the pharmaceutical compositioncontains no fish allergen. In another embodiment, the eicosapentaenoicacid or derivative thereof has a baseline peroxide value not greaterthan 5 meq/kg and upon storage of the composition at 25° C. and 60% RHfor a period of 6 months, the eicosapentaenoic acid or derivativethereof has a second peroxide value not greater than 8 meq/kg.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising eicosapentaenoic acid or a derivative thereof,octadecatetraenoic acid or a derivative thereof, nonaecapentaenoic acidor a derivative thereof, arachidonic acid or a derivative thereof,eicosatetraenoic acid or a derivative thereof, heneicosapentaenoate or aderivative thereof, and less than about 1%, by weight, of any fishallergens, if any. In another embodiment, the pharmaceutical compositioncontains less than about 0.05%, by weight, of fish allergens, if any. Inanother embodiment, the pharmaceutical composition contains less thanabout 0.005%, by weight, of fish allergens, if any. In anotherembodiment, the pharmaceutical composition contains less than about0.0005%, by weight, of fish allergens, if any. In another embodiment,the pharmaceutical composition contains less than about 0.00005%, byweight, of fish allergens, if any. In another embodiment, thepharmaceutical composition contains no fish allergen. In anotherembodiment, the eicosapentaenoic acid or derivative thereof has abaseline peroxide value not greater than 5 meq/kg and upon storage ofthe composition at 25° C. and 60% RH for a period of 6 months, theeicosapentaenoic acid or derivative thereof has a second peroxide valuenot greater than 8 meq/kg.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising at least 96%, by weight, of ethyleicosapentaenoate, from about 0.2% to about 0.5%, by weight, of ethyloctadecatetraenoate, from about 0.05% to about 0.25%. by weight, ofethyl nonaecapentaenoate, from about 0.2% to about 0.45%, by weight, ofethyl arachidonate, from about 0.3% to about 0.5%, by weight, of ethyleicosatetraenoate, from about 0.05% to about 0.32%, by weight, of ethylheneicosapentaenoate, less than about 0.05% by weight, of ethyl-DHA, ifany, and less than about 0.05% by weight, of fish allergens, if any.

In another embodiment, the present invention provides a method oftreating hypertriglyceridemia in a subject in need thereof, comprising:(a) identifying a subject having a fasting baseline triglyceride levelof about 500 mg/dl to about 2000 mg/dl and that is allergic to fish orseafood, and (b) administering to the subject a pharmaceuticalcomposition comprising about 1 g to about 4 g of a pharmaceuticalcomposition comprising at least about 96%, by weight, ethyleicosapentaenoate, wherein the composition contains substantially no DHAor derivative thereof and wherein upon administration the subject doesnot experience skin rash or experiences reduced incidence of skin rashcompared to placebo or to an equivalent milligram dose of a commerciallyavailable prescription omega-3 fatty acid composition (e.g. Lovaza). Inanother embodiment, the subject experiences no eructation or reducedeructation compared to placebo or to an equivalent milligram dose of acommercially available omega-3 fatty acid composition (e.g. Lovaza).

EXAMPLES

The following examples are for illustrative purposes only and are not tobe construed as limiting in any manner.

Example 1

A single center, double blind, randomized, parallel-group, placebocontrolled dose-ranging study of E-EPA in subjects with age-associatedimpairment (AAMI) was performed. The primary goal was to examine theeffect of ethyl-EPA versus placebo on cognitive performance in subjectswith AAMI as measure by the power of attention tasks in a computerizedtest batter over a period of 6 weeks. Secondary objectives were to:

(1) examine the effect of E-EPA versus placebo over 6 weeks on thefollowing tests in the computerized cognitive battery: Continuity ofattention tasks; Quality of working memory tasks; Quality of episodicmemory tasks; Speed of attention tasks;

(2) to assess the safety and tolerability of E-EPA versus placebo fromroutine clinical laboratory tests, adverse events (AE) monitoring andvital signs; and

(3) assess the potential dose-effect relationship of E-EPA on thecognitive endpoints by measurement of essential fatty acids in plasmaand red blood cell membranes. 94 subjects were randomized.

The study plan was to enroll 96 subjects who would be randomly allocatedto 1 of 4 possible treatment groups for 6 weeks, in a balanced blockdesign (24 subjects per group), as follows:

-   -   1. 1 g ethyl-EPA daily    -   2. 2 g ethyl-EPA daily    -   3. 4 g ethyl-EPA daily    -   4. Placebo (paraffin oil) daily

Ethyl-EPA was provided as 500 mg soft gel capsules providing ethyl-EPAof >96% purity, 0.25% to 0.38% by weight ethyl octadecatetraenoate,0.075% to 0.15% by weight ethyl nonaecapentaenoate, 0.25% to 0.35% byweight ethyl arachidonate, 0.3% to 0.4% by weight ethyleicosatetraenoate (ETA-E), 0.075% to 0.15% ethyl heneicosapentaenoateand 0.2% dl-□-tocopherol as an antioxidant. Matching placebo capsulescontained 467 g of liquid paraffin and 0.2% dl-□-tocopherol. The placebogroup was further randomized so that an equal number of subjects (8) wasallocated 1 g, 2 g or 4 g placebo. Study drug was taken twice daily(BID) as a divided dose (e.g. for the 1 g dose, 500 mg was given in themorning and a further 500 mg was given in the evening) with a lightsnack or meal.

The study consisted of a screening visit, a training visit and 4 studyvisits. At the screening visit, subjects' eligibility was determinedthrough cognitive tests (verbal paired associated learning [PAL]subscale, vocabulary subtest, Memory Assessment Clinics Questionnaire[MAC-Q], mini mental state evaluation [MMSE] and MINI [miniinternational neuropsychiatirc interview; sections 1 and 2 of Diagnosticand Statistical Manual of Mental Disorders, 4th Ed. (DSM-IV) plusdysthymia]), haematology, clinical chemistry and 12-leadelectrocardiogram (ECG). At the training visit, subjects were shown howto use the CDR computerized system. Subjects took study drug for 6 weeksand on Days 0, 14, 28 and 42, subjects underwent the CDR cognitive testbattery.

Inclusion Criteria

-   -   1. Written informed consent.    -   2. Male and female volunteers between 50 and 70 years of age.    -   3. Self-reported complaints of memory loss reflected in such        everyday problems as difficulty remembering names of individuals        following introduction, misplacing objects, difficulty        remembering multiple items to be purchased or multiple tasks to        be performed, problems remembering telephone numbers or postal        codes and difficulty recalling information quickly or following        distraction as determined by a score of 25 or higher on the        MAC-Q questionnaire. Onset of memory loss was to be described as        gradual without sudden worsening in recent months.    -   4. Possession of subjective and objective cognitive impairment        with a score of at least 1 standard deviation (SD) below that of        the mean for age-matched elderly population as determined by the        total score of between 13 and 20 from the PAL subset of the        Wechsler Memory Scale.    -   5. Evidence of adequate intellectual function as determined by a        scaled score of at least 9 (raw score of at least 32) on the        Vocabulary subtest of the Wechsler Adult Intelligence Scale        (WAIS).    -   6. Absence of dementia as determined by a score of 24 or higher        on the MMSE.    -   7. Non-smokers or ex-smokers for >3 months.    -   8. Was able to travel to the center and judged by the        Investigator as likely to be able to continue to travel for the        duration of the study and comply with the logistical aspects of        the study.    -   9. Body mass index (BMI) <29.5 kg/m².

Exclusion Criteria

-   -   1. Unlikely or unable to comply with investigational medication        dosing requirements.    -   2. Diagnosis of major depressive disorder, Alzheimer's or        vascular dementia as defined according to the MINI/DSM-IV Text        Revision (TR) criteria.    -   3. Past or current history of a neurological or psychiatric        disorder that could have affected cognitive function.    -   4. Past or current history of inflammatory gastrointestinal        disease such as Crohn's Disease or ulcerative colitis.    -   5. Constipation which required active treatment.    -   6. Current or previous history of cancer, excluding diagnosis of        basal cell carcinoma.    -   7. Any history or evidence of clinically significant cardiac        abnormality as measured by 12-lead ECG.    -   8. Any other medical condition or intercurrent illness not        adequately controlled, which, in the opinion of the        Investigator, may have put the subject at risk when        participating in the study or may have influenced the results of        the study or affected the subject's ability to take part in the        study.    -   9. Clinically significant abnormal screening laboratory results        (haematology, biochemistry) on screening or vital signs that        fell outside the normal range for this population, which in the        opinion of the Investigator affected the subject's suitability        for the study.    -   10. Any changes to prescribed medication for a medical condition        within 4 weeks of the baseline visit.    -   11. Omega-3 supplementation within 4 weeks of the baseline visit        or during the study treatment period.    -   12. Currently taking anticoagulants or daily dose of        aspirin >325 mg.    -   13. Cough or cold flu remedies containing opiates or        antihistamines, within 2 weeks of the baseline visit or during        the 6-week treatment period.    -   14. Known allergy to any ingredients of the study drug or        placebo.

Any subject could withdraw from the study at any time at their or theirlegal guardian's request, or at the discretion of the investigator, ifthe subjects continued inclusion was not in their best interest, or inthe event of a serious or unexpected AE. Every reasonable effort wasmade to document subject outcome and reasons for withdrawal. Any ongoingAEs were followed-up until the event had resolved, stabilized or wasotherwise explained. Subjects who were withdrawn were not replaced.Subjects were assigned unique identification numbers according to apre-determined randomization list generated by Catalent Pharma Solutionsand used in the drug packaging.

Study drug was administered orally BID as a divided dose with food, for6 weeks. Subjects were randomized to 1 of 6 possible treatment groups(Table 1).

TABLE 1 Treatment Groups Dosage Form (soft gel capsule) Group Dose (g)Study Drug Dosage Form (soft Group Dose (g) Study Drug gel capsule)Active 1 1 Ethyl-EPA 1 x 500 mg BID Active 2 2 Ethyl-EPA 2 x 500 mg BIDActive 3 4 Ethyl-EPA 4 x 500 mg BID Placebo 1 1 Paraffin oil 1 x 500 mgBID Placebo 2 2 Paraffin oil 2 x 500 mg BID Placebo 3 4 Paraffin oil 4 x500 mg BID BID = twice daily, ethyl-EPA = ethyl-eicosapentaenoic acid

Study drug was dispensed at Visits 3, 4 and 5; the maximum periodbetween Visit 3 and each subsequent visit was:

-   -   Visit 3 to Visit 4 (2 weeks±2 days from Visit 3).    -   Visit 3 to Visit 5 (4 weeks±2 days from Visit 3).    -   Visit 3 to Visit 6 (6 weeks±2 days from Visit 3).

All treatment packs were identical in appearance, in order to maintainsubject and investigator blind throughout the study. The investigator,Sponsor/clinical research organization personnel and subjects remainedblinded throughout this study. The investigator was permitted toun-blind individual subjects if it was considered medically imperative.The process for breaking the blind is outlined below.

Omega-3 supplements had to be discontinued at least 4 weeks prior to thebaseline visit (Visit 3). Cough and influenza remedies containingopiates or antihistamines had to be discontinued 2 weeks prior to thebaseline visit (Visit 3) and were not permitted for the duration of thestudy.

Existing medication had to have been stable for 4 weeks prior to thebaseline visit (Visit 3) and the dose maintained for the duration of thestudy. Where a dose change was absolutely necessary this was recorded inthe electronic case report form (eCRF).

Subjects who required anticoagulant medication during the study were tobe withdrawn. Psychological counseling or therapy was not permitted forthe duration of the study, as these could have interfered with theoutcome of the study. Unused study drug was returned to the study site.Subjects who used less than 80% of the prescribed dose were considerednon-compliant.

At screening cognitive testing and suitability for the study wereassessed using the Verbal Paired Associates 1 (Wechsler Memory Scale),Vocabulary Subtest of the WAIS, MAC-Q, MMSE and MINI (DSM-IV Sections 1and 2 plus Dysthymia).

A selection of tasks from the CDR computerized cognitive assessmentsystem were administered (Appendix 8 of protocol) at Visit 2 (trainingvisit), Visit 3 (baseline), Visit 4 (Day 14), Visit 5 (Day 28) and Visit6 (Day 42). Parallel forms of the tests were presented at each testingsession. All tasks were computer-controlled, the information presentedon high resolution monitors, and the responses recorded via a responsemodel containing 2 buttons 1 marked ‘no’ the other ‘yes’. Five CDRcomposite scores were used as the primary/secondary outcome variables.

The task titles were:

Word Presentation Numeric Working Memory Immediate Word Recall DelayedWord Recall Picture Presentation Word Recognition Simple Reaction TimePicture Recognition Digit Vigilance Bond-Lader Visual Choice ReactionTime Analogue Scales of Mood Spatial Working Memory and AlertnessScreen, Using the Computer Mouse

To ensure consistency of approach, full training on the cognitive testsand CDR test battery was provided to study site staff and studysubjects. The results of each variable were automatically recorded usingthe machine interface developed by CDR.

An AE was defined as any untoward medical occurrence temporallyassociated with the use of a medicinal product whether or not consideredrelated to the medicinal product.

The investigator was responsible for the detection and documentation ofAEs. At each visit the subject was asked about AEs by means ofnon-leading questions. AEs were recorded from the time a subjectprovided a written informed consent and deemed eligible to participateuntil completion of the treatment period. AEs ongoing at the end of thetreatment period were followed until resolution or return to baseline ornormal value or if the event was considered unrelated to study drug.

A serious adverse event (SAE) was defined as any AE at any dose that:

-   -   resulted in death;    -   was life-threatening;    -   required hospitalization or prolongation of existing        hospitalization;    -   resulted in disability or incapacity, or    -   resulted in a congenital anomaly/birth defect.

Other events were considered SAEs if they jeopardized the subject orrequired medical or surgical intervention to prevent one of the outcomeslisted above.

Regardless of the above criteria, any AE that the Sponsor orinvestigator considered serious was to have been immediately reported asa SAE. Any death or SAE experienced by the patient while receiving orwithin 30 days of last dose of Investigational Medicinal Product must bepromptly reported (within 24 hours of learning of the event) topharmacovigilance. All AEs (including SAEs) are to be accuratelyrecorded on the adverse event page of the subject's eCRF, beginning fromfirst administration of Investigational Medicinal Product until 30 daysafter the last dose.

Blood samples for the laboratory assessments for haematology (a 5 mLblood sample) and clinical chemistry (a 10 mL blood sample) listed inTable 2, were collected at the screening visit (Visit 1). Samples wereprocessed and analyzed by Simbec Laboratories Ltd.

TABLE 2 Laboratory Assessments Clinical Chemistry Haematology Sodium Redblood cell count Potassium White blood cell count Bicarbonate Meancorpuscular volume Urea Mean corpuscular haemoglobin Creatinine Meancorpuscular haemoglobin concentration Total bilirubin HaemoglobinAspartate aminotransferase Platelet count Alanine aminotransferaseNeutrophils Gamma glutamyl transferase Lymphocytes Total proteinMonocytes Albumin Glucose Basophils

Pharmacodynamic: Essential Fatty Acid (EFA) Measurements

Blood samples (10 mL) were collected at Visit 1 (screening) and atVisits 4, 5 and 6. Analysis was performed by MSR Lipid Analysis,Scottish Crop Research Institute, Dundee, UK. The screening sample actedas baseline for the EFA measurements.

Lipid was extracted from plasma, serum and RBC suspensions and convertedinto fatty acid methyl esters which were analyzed by gas chromatographyto give fatty acid profiles as micrograms fatty acid per gram of sample(μgFA/g) and normalized area percent. The CDR computerized system hasbeen used to measure the effects of pharmaceuticals on cognitivefunction in a variety of clinical trials. Efficacy was assessed by abattery of cognition tests designed by CDR. Safety data were analyzed byQuanticate.

Populations analyzed included:

-   -   Intent to Treat (ITT) Population: All randomized subjects with        at least 1 visit post-baseline were included in this population,        regardless of treatment actually received.    -   Per Protocol Population (PP): All randomized subjects that        completed the study, excluding significant protocol deviators,        were defined as the Safety PP population. An Efficacy PP        population was based on the Efficacy completers. The intercept        of the Safety and Efficacy PP populations defined the Study PP        Population.    -   Safety Population: All randomized subjects that received at        least 1 dose of study medication.

Summary statistics were provided for the ITT and Study PP Populationsseparately for all composite scores, major and supportive variables.Summary statistics were performed for both the unadjusted and differencefrom baseline data (i.e. the difference from the time matched pre-doseassessments on Day 0). Summary statistics were calculated by treatment,day and time-point. The summary statistics comprised n, mean, median,SD, standard error of mean (SEM), minimum and maximum values.

Difference from baseline data for each major variable was evaluated byan Analysis of Covariance (ANCOVA) using SAS® PROC MIXED Version 8.2.

Fixed effects for treatment, day, time point, treatment by day,treatment by time point, treatment by day by time-point were fitted.Subject within treatment was fitted as a repeated effect using therepeated statement. The compound symmetry covariance structure was used.Subjects' time-matched predose assessments on Day 0 were used as acovariate in the analysis. Least squares means (LS means) werecalculated for treatment by day, treatment by time-point and treatmentby day by time-point interaction. This formal analysis was conducted forthe ITT and Study PP Populations separately.

Safety evaluations were based on the safety population. Safety andtolerability were assessed in terms of AEs, vital signs, 12-lead ECG,clinical laboratory data, medical history, and study drug compliance.Safety and tolerability data were presented by treatment group. Allsafety data were listed individually by subject.

RBC and plasma EFA data were collected at baseline, Day 14, 28 and 42and summarized by visit for each treatment group. Change from baselineand percent change from baseline were also summarized. ANCOVA comparisonof ethyl-EPA dose groups and ethyl-EPA versus placebo was performed.

The sample size calculation was based on Power of Attention.Ispronicline (50 mg), a neuronal nicotinic acetylcholine receptorpartial agonist, in subjects with AAMI on Day 21 of repeated dosing inan earlier study showed a benefit of 61 msec (50 mg mean=−32.54,SD=61.22; placebo mean=28.25, SD=49.64) to Power of Attention. Using apooled SD, a sample size of 15 subjects per treatment arm was consideredsufficient to detect a difference of 61 msec, with 80% power and 5%significance level (no adjustment for multiple testing). As there was noprior experience with the compound or mechanism of action with thesecognitive measures, a sample size of 24 subjects per treatment arm waschosen as sufficient to allow for early withdrawals.

There were no changes to the conduct of the study. The following changeswere made to the planned analyses: The equation to calculate Speed ofMemory was changed to SPEEDMEM (speed of memory)=SPMRT (spatial workingmemory speed)+NWMRT (numeric working speed)+DRECRT (word recognitionspeed)+DPICRT (picture recognition speed).

-   -   Subject's time-matched pre-dose assessments on Day 0 were used        as a covariate in the analysis.    -   Day 0 was removed from Day values in the list of ANCOVA variable        values. Covariate=Baseline was changed to Covariate=Time matched        predose assessments on Day 0 in the list of ANCOVA variable        values.    -   Day by Time-point was added to the list of model effects in SAS®        code for ANCOVA model.    -   F Tests table and Treatment Effects table were added to list of        ANCOVA summary tables.    -   ANCOVA summary tables were renumbered to follow on from ANCOVA        raw outputs.    -   Figures were included for Treatment, Treatment by Day, Treatment        by Time-point, Treatment by Day by Time-point effects for ANCOVA        LS means.    -   Figures were added for ANCOVA LS means differences to placebo        (95% confidence interval [CI]).    -   A post-hoc analysis was performed which compared the individual        placebo groups (1 g, 2 g and 4 g paraffin oil) with the        corresponding ethyl-EPA dose rather than to a pooled placebo        group.

Ninety-one subjects completed the study, three subjects discontinued; 2subjects from the ethyl-EPA 2 g treatment group (1 subject due to an SAEconsidered unrelated to the study drug and 1 due to a protocol violationand 1 subject from the placebo 2 g group due to an AE.

For Power of Attention, there was no statistically significant effect oftreatment, nor any treatment by day, treatment by time-point ortreatment by day by time-point interactions. There was no LS meandifference between active treatment and placebo at any time-point. ForChoice Reaction Time there were statistically significant benefits forethyl-EPA 1 g and 2 g on Day 28, and some trends for benefit for 1 and 4g ethyl-EPA on Day 42, versus placebo; however, no cleartreatment-related pattern was observed.

Continuity of Attention did not show a difference between placebo andethyl-EPA, except for an overall decrease for 2 g ethyl-EPA that wasonly visible in the ITT population. The subtask Digit Vigilance TargetsDetected showed isolated decreases for active treatment versus placebo,but there was no obvious treatment-related pattern.

Quality of Working Memory was the only composite score that showed astatistically significant treatment by day interaction in the F-ratio.However, there were only isolated statistically significant decreasesfor ethyl-EPA 1 g and 2 g versus placebo on Days 14 and 28, and thesewere most likely to be due to chance and not treatment related.

Quality of Episodic Secondary Memory showed statistically significantdecreases for ethyl-EPA versus placebo at various time-points. However,it seems unlikely to be an effect of active treatment as the unadjusteddata showed pre-existing differences between the treatment groups thatwas most notable on Day 0 in the first assessment session. In differencefrom Baseline data that were calculated prior to ANCOVA analysis, thesedifferences were no longer apparent. This suggests that the ANCOVA modelfitted a strong negative correlation with the baseline values. This isoften the case when the variability within subjects overlaps thevariability between subjects.

Speed of Memory and the subtasks Spatial and Numeric Working MemorySpeeds and Word and Picture Recognition Speed showed no differences inperformance, in the F-ratio statistics, between Ethyl-EPA and placebo.

For Self-rated Alertness, there was no apparent difference in ratingsbetween ethyl-EPA and placebo. There were isolated decreases in ratingsfor active treatment versus placebo that were unlikely to be compoundrelated.

Self-rated Contentment showed statistically significant decreases inratings for ethyl-EPA 2 g on Day 28. However, these individual decreaseswere not statistically significant. It is unlikely that this was atreatment-related effect as it was restricted to a single day and noother dose level showed a similar pattern on any other day. ForSelf-rated Calmness there was no difference in ratings between activetreatment and placebo.

When the results of each ethyl-EPA dose and their corresponding placebowere compared (post-hoc analysis), it appeared that ethyl-EPA 4 gimproved the subjects' reaction times in the attention tasks (Power ofAttention, Simple Reaction Time and Choice Reaction Time). This was seenmost clearly for Choice Reaction Time, where a pattern of gradualimprovement over the assessment day for 4 g was seen. It is possiblethat a longer period of administration would clarify the effects ofethyl—EPA on these parameters.

EPA (shown in Table 3), DPAn-3 and EPA/AA ratio (data not shown) plasmaand RBC values increased substantially from baseline to Day 42 for theAMR-101 1, 2, and 4 g treatment groups. AA, DHA and DGLA valuesdecreased substantially from baseline (data not shown).

TABLE 3 Mean (SD) EPA (Plasma and RBC (μg/g)) Change from Baseline.Ethyl-EPA Placebo 1 g (N = 23) 2 g (N = 24) 4 g (N = 24) 1 g (N = 7) 2 g(N = 8) 4 g (N = 8) Plasma Baseline 48.3 (31.03)  44.9 (25.01)  49.1(17.23) 47.5 (26.41) 42.1 (16.18) 42.5 (11.86) Day 14 61.2 (26.61) 124.6(42.25) 207.7 (57.05)  1.6 (24.69 −1.2 (19.82) 21.9 (32.91) Day 28 60.3(36.03) 142.2 (46.23) 215.2 (58.68)  6.5 (15.46)  1.6 (13.64)  1.3(14.03) Day 42 62.0 (39.43) 133.4 (43.34) 204.6 (80.69) 11.9 (26.34) 0.4 (21.18)  4.4 (23.32) RBC Baseline 19.8 (10.85) 18.9 (8.91) 19.8(5.28) 20.4 (5.77)  19.3 (6.58)  17.2 (4.94)  Day 14 12.3 (7.39)  26.9(9.15)  39.5 (13.16) −0.5 (6.32)  0.0 (7.17) 2.6 (6.73) Day 28 14.5(10.47)  32.9 (10.11)  50.2 (15.82) 1.5 (4.16) 0.0 (7.06  0.6 (4.42) Day42 17.6 (11.89)  38.3 (12.46)  52.5 (20.56) −0.2 (5.90)  1.0 (8.01) −0.2(6.97) 

As can be seen in Table 3, at the 2 g per day AMR101 dose, plasma EPAlevels increased 297% after 42 days and at the 4 g per day AMR101 dose,plasma EPA levels increased by 417% compared to baseline or placebocontrol.

Grimsgaard et al. previously published an article describing serumphospholipid levels at baseline and after 7 weeks of supplementationwith 4 g per day of 90% ethyl-DHA, 4 g per day of 95% ethyl-EPA withsome DHA present, or corn oil. Am. J. Clin. Nutr. 1997; 66:649-59(1997). The complete profile of additional fatty acids and ingredientspresent in these compositions is unknown. After supplementation over aperiod of 7 weeks, subjects exhibited only a 297% increase in serumphospholipid EPA compared to the increase of 417% shown above with aninventive composition. A comparison of other chances in plasma/serumfatty acids is shown in Table 4.

TABLE 4 Percent Fatty Acid Change from Baseline After Administration of4 g Dose Fatty Acid Grimsgaard AMR101 EPA  +297%  +417% AA −18.5% −21.9%DHA −15.20%  −17.5% DPA  +130%  +147% DGLA −30.5% −39.4%

Furthermore, in the Japanese Eicosapentaenoic Acid (EPA) LipidIntervention Study (JELIS), Yokoyama et al. reported that they followedover 18,000 patients randomly assigned to received either 1800 mg of EPAcomposition (Epadel) with statin, or statin only with a 5-yearfollow-up. Lancet 2007; 369: 1090-98. After 5 years of treatment,subjects exhibited an increase in plasma EPA of only 70% (from baselineof 93 mg/L to 169 mg/L).

FIGS. 1 and 2 and show a comparison of the change in plasma/serum EPAlevels observed with AMR101 treatment in the current study compared tothose observed with different EPA compositions in the JELIS study and byGrimsgaard. As will be noted, at ˜2 g per day, AMR101 achieved muchgreater plasma EPA increase compared to baseline (˜4-fold) after just 6weeks than the JELIS study observed (<2-fold) after 5 years oftreatment. Moreover, at the 4 g per day dose, AMR101 treatment for 6weeks achieved much higher (>250 μg/g) plasma EPA levels than reportedby Grimsgaard after 7 weeks of treatment (87.66 μg/g serum). Overall,the 4 g per day dose of AMR101 resulted in a greater than 5-foldincrease in plasma EPA over baseline while the 4 g per day dose ofGrimsgaard's composition resulted in less than a 3-fold increase inserum EPA. These results were unexpected.

Example 2

A multi-center, randomized, double-blind, placebo-controlled trial wasconducted in North America to determine whether 1 gram twice daily ofEPA for 6 months improves motor performance in Huntington's patients. Apost-hoc analysis was performed to evaluate the effect of EPA onnon-fasting triacylglycerols.

Study of the effects of ethyl-EPA on the progression of HuntingtonDisease enrolled study participants at 41 sites in Canada and the UnitedStates. Based on the results of the earlier study, the study entrycriteria were designed to enrich the participation of individuals withHuntington disease with a CAG repeat less than 45, without requiringgenetic testing to reveal the length of expansions to researchparticipants or investigators. To participate in the study, individualshad to have the clinical features of HD and either a confirmatory familyhistory or a known CAG expansion. Eligibility criteria included aminimum age of 35, a total functional capacity of at least 7, minimaldystonia (not exceeding 2 on the UHDRS in either the trunk orextremities), minimal bradykinesia (not exceeding 2 on the UHDRS itemfor bradykinesia), the use of adequate birth control, the ability totake oral medications, and the willingness and ability to comply withstudy requirements. Individuals were not eligible to participate if,within 60 days of the baseline visit, they had used omega-3 fatty acidsupplements, tetrabenazine or reserpine, high or variable doses of oralanti-psychotic medications (e.g., haloperidol), steroids other thantopical preparations, high dose selenium supplements, lithium, highdoses of benzodiazepines, anti-coagulation medication (e.g., coumadin),high doses (greater than 325 mg per day) of aspirin, unstable does ofNMDA receptor antagonists (e.g., memantine), unstable doses ofanti-epileptic medications, or if they had participated in otherinvestigational drug studies. Additional exclusion criteria were the useof depot neuroleptics within 6 months of the baseline visit, a historyof tardive dyskinesia, unstable medical or psychiatric illness, majordepression (defined as a score greater than 20 on the Beck DepressionInventory II), suicidal ideation, clinically significant substance abusewithin 12 months of the baseline visit, women who were pregnant orlactating, known allergy to ethyl-EPA or placebo, or previousparticipation in an investigational study of EPA.

This was a randomized, double-blind, placebo-controlled, parallel groupstudy of EPA (1 gram twice/day). The institutional review board at eachparticipating site approved the research plan and consent documents.Eligible study participants provided written consent. At the baselinevisit, participants were randomized according to a block-balancedcomputer-generated randomization plan that was stratified by site andgenerated by the Biostatistics Center at the University of Rochester.Individuals were randomized in a 1:1 ratio to receive either active drug(n=158) in the form of two 500 mg capsules of AMR101 orally or placebo(n=154) in the form of two 500 mg capsules containing light paraffin oiland 0.2% dl-alpha-tocopherol twice daily orally for 6 months. After 6months, all TREND-HD participants were treated with AMR101 for 6 monthsin an open-label fashion. Only data from the first 6 months were used toevaluate the effects of AMR101 on lipids.

The outcome measure of this study was the change in non-fastingtriacylglycerol (TG) levels in those on AMR101 compared to those onplacebo.

Safety was assessed at all study visits, including evaluation andassessment of adverse events and serious adverse events and review ofclinical laboratory tests (complete blood count, serum chemistry, andurine pregnancy tests). The safety of research participants wasmonitored in a blinded manner by a medical monitor from both the sponsorand from the Huntington Study Group. In addition, an independent SafetyMonitoring Committee that had access to treatment assignments reviewedsafety data throughout the study to determine if any modifications wereneeded to the trial's conduct.

Changes in lipid levels were compared using an analysis of covariance(ANCOVA) with treatment group as the factor of interest, site as astratification factor, and baseline value as a covariate. Allindividuals who received study medication were included in the safetyanalysis. For each type of adverse event, the treatment groups werecompared regarding the occurrence of at least one event using Fisher'sexact test. Continuous measures of safety such as laboratory testresults and vital signs were analyzed using methods similar to thosedescribed above for the primary outcome variable (ANCOVA). Nocorrections were made for multiple comparisons in evaluating safetydata.

One hundred forty-five subjects on AMR101 (92% of those assigned) and141 of those on placebo (92% of those assigned) had red blood cellcontent of EPA determined at baseline and 6 months. Baseline red bloodcell content of 20:5n3 (EPA) increased significantly after 6 months inthose on AMR101 (from a mean of 0.52% to 3.07%) but decreased in thoseon placebo (from a mean of 0.61% to 0.55%); p<0.0001). After 6 months,individuals taking AMR101 had a 26 mg/dL decrease in TGs from a baselineof 171 compared to a decrease of 11 mg/dL from a baseline of 187 mg/dLin those on placebo; p=0.007. Total cholesterol was reducedsignificantly more in those taking AMR101 (9.5 mg/dL) from a baseline of204 mg/dL than in those taking placebo (2.5 mg/dL) from a baseline of208 mg/dL; p=0.009. Lipid and Motor Scoer data are shown in Tables 5 and6, respectively.

TABLE 5 Motor Score Results. Total motor score 4 of the All StudyParticipants Study Participants with CAG < 45 Unified Huntington's n =316 n = 221 Disease Rating Scale Ethyl-EPA Placebo p-value Ethyl-EPAPlacebo p-value At baseline [mean (SD)] 25.2 (8.3) 23.9 (8.1) 0.16 24.9(8.3) 23.4 (7.7) 0.18 Change in total motor score 4 0.2 1.0 0.20 0.0 0.30.70 at 6 months (mean) Change in total motor score 4 0.0 2.0 0.02 −1.21.6 0.004 at 12 months (mean)

TABLE 6 Lipid Parameter Results. p- Lipoprotein Variable Ethyl-EPAPlacebo value Baseline triglycerides (mean 171 ± 108 187 ± 139 0.27mg/dL ± SD) Baseline total cholesterol  204 ± 41.4  208 ± 40.6 0.42(mean mg/dL ± SD) Change in triglycerides after −25.8 ± 89.1  −11.1 ±105.2 .007 6 months (mean mg/dL ± SD) Change in total cholesterol after−9.5 ± 28.6 −2.5 ± 24.7 .009 6 months (mean mg/dL ± SD) Change intriglycerides after −17.7 ± 86.7  −40.0 ± 126.0 0.66 12 months (meanmg/dL ± SD) Change in total cholesterol after −5.6 ± 25.5 −6.9 ± 34.50.95 12 months (mean mg/dL ± SD)

By comparison with these data for AMR101, Grimsgaard reported a decrease(from baseline) of only 12% in serum triglycerides in the EPA groupafter 7 weeks of treatment. Furthermore, addition of the Epadel EPAcomposition to existing statin therapy in the JELIS study resulted inonly a 9% reduction in triglycerides after 5 years of treatment.

Example 3

A study was performed to evaluate and compare the content of Epadelcapsules with AMR101 capsules. Six capsules of each composition wereselected for analysis by gas chromatography. Averages of the sixcapsules for each of the two compositions are shown in Table 7.

TABLE 7 Measured and Identified Components of AMR101 and Epadel. AMR101Epadel Component Amount (% w/w) Ethyl-EPA 96.3 94.5 ODTA-E 0.25 0.09Impurity 3 ND 0.06 NDPA-E 0.11 0.11 Impurity 4 0.08 0.07 AA-E 0.30 0.06ETA-E 0.38 0.11 Isomer A 0.08 0.23 Isomer D, E 0.11 0.62 HPA-E 0.11 0.06ND = w/w % less than 0.05%

Example 4

A phase I, multiple dose pharmacokinetic study in healthy malevolunteers was carried out at a single center. Twenty-four subjects weredivided into two treatment groups of 12 subjects each (groups A and B).Both groups received the same total daily dose of AMR101 but the dosingregiments were different. All subjects received a single oral dose of 2g AMR101 on Day 1. Treatment Group A received 28 continuous once dailydoses of 2 g AMR101. Treatment Group B received 27 continuous twicedaily doses of 1 g AMR101 and a single dose of 2 g of AMR101 on day 30.

Levels of EPA and other essential fatty acids were determined in plasmaand red blood cells. Blood samples for pharmacokinetic analysis weretaken at the following time points for Treatment groups A and B:

Days 1 and 30: Pre-dose, 1, 2, 3, 4, 5, 6, 8, 20, 12, 24, 36 and 48 h.post-dose;

Days 9, 16, 23: pre morning dose;

Days 37, 44, 58: post last dose.

A first Interim Report presents the following pharmacokinetic resultsfor Treatment Group B:

Plasma—Day 1 (Pre-dose, 1, 2, 3, 4, 5, 6, 8, 20, 12, 24, 36 and 48 hpost-dose);

Red cell—Day 1 (Pre-dose and 36 h), Day 30 (1 h post-dose), Day 37, Day44, Day 58.

Using a corrected value obtained by subtracting the pre-administrationconcentration from the concentrations at each sampling, a single oraldose of 2 g of AMR101 resulted in a rapid rise in plasma lipid EPA.Maximum values were observed at 5 hours post-administration with EPAlevels remaining above baseline at 48 hours post-administration. Thehalf-life of removal of EPA from plasma lipids was 87±65 h (non-baselinesubtracted) and 42±31 h (baseline subtracted). Summary pharmacokineticdata are shown in Table 8.

TABLE 8 Non-Compartmental Analysis - Arithmetic Mean and SD. Mean VoD atTerminal Residence Oral Terminal VoD at Max Drug Half-Life Time (h)Clearance Phase Steady State Conc. (mg/ml) Tmax (h) Unadjusted 86.6126.6 0.381 37.0 37.8 78.3 4.64 SD 65.4 93.3 0.202 13.2 13.5 33.7 0.92Baseline 42.2 63.6 1.27 58.8 62.8 55.5 4.64 Subtracted 0.021 30.9 43.10.83 23.9 25.7 28.2 0.92

In the Per Protocol population oral administration of AMR101 resulted inRBC EPA levels increasing from a mean value of 190.4 mg/g before dosingon Day 1 to 40.3 mg/g one hour following the final dose on Day 30.

Example 5

A multi-center, placebo-controlled randomized, double-blind, 12-weekstudy with an open-label extension was performed to evaluate theefficacy and safety of AMR101 in patients with fasting triglyceridelevels ≥500 mg/dL. The primary objective of the study was to determinethe efficacy of AMR101 2 g daily and 4 g daily, compared to placebo, inlowering fasting TG levels in patients with fasting TG levels ≥500 mg/dLand ≤1500 mg/dL (≥5.65 mmol/L and ≤16.94 mmol/L).

The secondary objectives of this study was the following:

-   1. To determine the safety and tolerability of AMR101 2 g daily and    4 g daily;-   2. To determine the effect of AMR101 on lipid and apolipoprotein    profiles;-   3. To determine the effect of AMR101 on low-density lipoprotein    (LDL) particle number and size;-   4. To determine the effect of AMR101 on oxidized LDL;-   5. To determine the effect of AMR101 on fasting plasma glucose (FPG)    and hemoglobin A_(1c) (HbA_(1c));-   6. To determine the effect of AMR101 on insulin resistance;-   7. To determine the effect of AMR101 on high-sensitivity C-reactive    protein (hsCRP);-   8. To determine the effects of AMR101 2 g daily and 4 g daily on the    incorporation of fatty acids into red blood cell membranes and into    plasma phospholipids;-   9. To explore the relationship between baseline fasting TG levels    and the reduction in fasting TG levels; and-   10. To explore the relationship between an increase in red blood    cell membrane eicosapentaenoic acid (EPA) concentrations and the    reduction in fasting TG levels.

The population for this study is men and women (women of childbearingpotential will need to be on contraception or practice abstinence) >18years of age with a body mass index ≤45 kg/m² who are not onlipid-altering therapy or are currently on lipid-altering therapy.Patients currently on statin therapy (with or without ezetimibe) wereevaluated by the investigator as to whether this therapy can be safelydiscontinued at screening, or if it should be continued. If statintherapy (with or without ezetimibe) is to be continued, dose(s) werestable for ≥4 weeks prior to randomization. Patients taking non-statin,lipid-altering medications (niacin >200 mg/day, fibrates, fish oil,other products containing omega-3 fatty acids, or other herbal productsor dietary supplements with potential lipid-altering effects), eitheralone or in combination with statin therapy (with or without ezetimibe),were required to safely discontinue non-statin, lipid-altering therapyat screening.

Approximately 240 patients were randomized at approximately 50 centersin North America, South America, Central America, Europe, India, andSouth Africa. The Phase 3, multi-center study consisted of 3 studyperiods: (1) A 6- to 8-week screening period that includes a diet andlifestyle stabilization and washout period and a TG qualifying period;(2) A 12-week, double-blind, randomized, placebo-controlled treatmentperiod; and (3) A 40-week, open-label, extension period.

During the screening period and double-blind treatment period, allvisits were within ±3 days of the scheduled time. During the open-labelextension period, all visits were within ±7 days of the scheduled time.The screening period included a 4- or 6-week diet and lifestylestabilization period and washout period followed by a 2-week TGqualifying period.

The screening visit (Visit 1) occurred for all patients at either 6weeks (for patients not on lipid-altering therapy at screening or forpatients who did not need to discontinue their current lipid-alteringtherapy) or 8 weeks (for patients who required washout of their currentlipid-altering therapy at screening) before randomization, as follows:

Patients who did not require a washout: The screening visit occurred atVisit 1 (Week −6). Eligible patients entered a 4-week diet and lifestylestabilization period. At the screening visit, all patients receivedcounseling regarding the importance of the National CholesterolEducation Program (NCEP) Therapeutic Lifestyle Changes (TLC) diet andreceived instructions on how to follow this diet. Patients who requireda washout: The screening visit occurred at Visit 1 (Week −8). Eligiblepatients began a 6-week washout period at the screening visit. Patientsreceived counseling regarding the NCEP TLC diet and receivedinstructions on how to follow this diet. Site personnel contactedpatients who did not qualify for participation based on screeninglaboratory test results to instruct them to resume their priorlipid-altering medications.

At the end of the 4-week diet and lifestyle stabilization period or the6-week diet and stabilization and washout period, eligible patientsentered the 2-week TG qualifying period and had their fasting TG levelmeasured at Visit 2 (Week −2) and Visit 3 (Week −1). Eligible patientshad an average fasting TG level ≥500 mg/dL and ≤1500 mg/dL (≥5.65 mmol/Land ≤16.94 mmol/L) to enter the 12-week double-blind treatment period.The TG level for qualification was based on the average (arithmeticmean) of the Visit 2 (Week −2) and Visit 3 (Week −1) values. If apatient's average TG level from Visit 2 and Visit 3 fell outside therequired range for entry into the study, an additional sample forfasting TG measurement was collected 1 week later at Visit 3.1. If athird sample was collected at Visit 3.1, entry into the study was basedon the average (arithmetic mean) of the values from Visit 3 and Visit3.1.

After confirmation of qualifying fasting TG values, eligible patientsentered a 12-week, randomized, double-blind treatment period. At Visit 4(Week 0), patients were randomly assigned to 1 of the followingtreatment groups:

-   -   AMR101 2 g daily,    -   AMR101 4 g daily, or    -   Placebo.

During the double-blind treatment period, patients returned to the siteat Visit 5 (Week 4), Visit 6 (Week 11), and Visit 7 (Week 12) forefficacy and safety evaluations.

Patients who completed the 12-week double-blind treatment period wereeligible to enter a 40-week, open-label, extension period at Visit 7(Week 12). All patients received open-label AMR101 4 g daily. From Visit8 (Week 16) until the end of the study, changes to the lipid-alteringregimen were permitted (e.g., initiating or raising the dose of statinor adding non-statin, lipid-altering medications to the regimen), asguided by standard practice and prescribing information. After Visit 8(Week 16), patients returned to the site every 12 weeks until the lastvisit at Visit 11 (Week 52).

Eligible patients were randomly assigned at Visit 4 (Week 0) to receiveorally AMR101 2 g daily, AMR101 4 g daily, or placebo for the 12-weekdouble-blind treatment period. AMR101 was provided in 1 g liquid-filled,oblong, gelatin capsules. The matching placebo capsule was filled withlight liquid paraffin and contained 0 g of AMR101. During thedouble-blind treatment period, patients took 2 capsules (AMR101 ormatching placebo) in the morning and 2 in the evening for a total of 4capsules per day. Patients in the AMR101 2 g/day treatment groupreceived 1 AMR101 1 g capsule and 1 matching placebo capsule in themorning and in the evening. Patients in the AMR101 4 g/day treatmentgroup received 2 AMR101 1 g capsules in the morning and evening.

Patients in the placebo group received 2 matching placebo capsules inthe morning and evening. During the extension period, patients receivedopen-label AMR101 4 g daily. Patients took 2 AMR101 1 g capsules in themorning and 2 in the evening.

The primary efficacy variable for the double-blind treatment period waspercent change in TG from baseline to Week 12 endpoint. The secondaryefficacy variables for the double-blind treatment period included thefollowing:

-   -   Percent changes in total cholesterol (TC), high-density        lipoprotein cholesterol (HDL-C), calculated low-density        lipoprotein cholesterol (LDL-C), calculated non-high-density        lipoprotein cholesterol (non-HDL-C), and very low-density        lipoprotein cholesterol (VLDL-C) from baseline to Week 12        endpoint;    -   Percent change in very low-density lipoprotein TG from baseline        to Week 12;    -   Percent changes in apolipoprotein A-I (apo A-I), apolipoprotein        B (apo B), and apo A-I/apo B ratio from baseline to Week 12;    -   Percent changes in lipoprotein(a) from baseline to Week 12        (selected sites only);    -   Percent changes in LDL particle number and size, measured by        nuclear magnetic resonance, from baseline to Week 12 (selected        sites only);    -   Percent change in remnant-like particle cholesterol from        baseline to Week 12 (selected sites only);    -   Percent change in oxidized LDL from baseline to Week 12        (selected sites only);    -   Changes in FPG and HbA_(1c) from baseline to Week 12;    -   Change in insulin resistance, as assessed by the homeostasis        model index insulin resistance, from baseline to Week 12;    -   Percent change in lipoprotein associated phospholipase A2 from        baseline to Week 12 (selected sites only);    -   Change in intracellular adhesion molecule-1 from baseline to        Week 12 (selected sites only);    -   Change in interleukin-6 from baseline to Week 12 (selected sites        only);    -   Change in plasminogen activator inhibitor-1 from baseline to        Week 12 (selected sites only);    -   Change in hsCRP from baseline to Week 12 (selected sites only);    -   Change in serum phospholipid EPA content from baseline to Week        12;    -   Change in red blood cell membrane EPA content from baseline to        Week 12; and    -   Change in serum phospholipid and red blood cell membrane content        in the following fatty acids from baseline to Week 12:        docosapentaenoic acid, docosahexaenoic acid, arachidonic acid,        palmitic acid, stearic acid, and oleic acid.

The efficacy variable for the open-label extension period was percentchange in fasting TG from extension baseline to end of treatment. Safetyassessments included adverse events, clinical laboratory measurements(chemistry, hematology, and urinalysis), 12-lead electrocardiograms(ECGs), vital signs, and physical examinations

For TG, TC, HDL-C, calculated LDL-C, calculated non-HDL-C, and VLDL-C,baseline was defined as the average of Visit 4 (Week 0) and thepreceding lipid qualifying visit (either Visit 3 [Week −1] or if itoccurs, Visit 3.1) measurements. Baseline for all other efficacyparameters was the Visit 4 (Week 0) measurement.

For TC, HDL-C, calculated LDL-C, calculated non-HDL-C, and VLDL-C, Week12 endpoint was defined as the average of Visit 6 (Week 11) and Visit 7(Week 12) measurements. Week 12 endpoint for all other efficacyparameters was the Visit 7 (Week 12) measurement.

The primary efficacy analysis was performed using a 2-way analysis ofcovariance (ANCOVA) model with treatment as a factor and baseline TGvalue as a covariate. The least-squares mean, standard error, and2-tailed 95% confidence interval for each treatment group and for eachcomparison was estimated. The same 2-way ANCOVA model will be used forthe analysis of secondary efficacy variables.

The primary analysis was repeated for the per-protocol population toconfirm the robustness of the results for the intent-to-treatpopulation.

The primary efficacy variable was the percent change in fasting TGlevels from baseline to Week 12. A sample size of 69 completed patientsper treatment group provided ≥90% power to detect a difference of 30%between AMR101 and placebo in percent change from baseline in fasting TGlevels, assuming a standard deviation of 45% in TG measurements and asignificance level of p<0.01. To accommodate a 15% drop-out rate fromrandomization to completion of the double-blind treatment period, atotal of 240 randomized patients was planned (80 patients per treatmentgroup).

Top line results of the study are shown in FIGS. 3 and 4. Adverse eventsare shown in Table 9 by comparison with placebo and those experienced bysubjects taking commercially available mixture of E-EPA and E-DHA(Lovaza).

AMR101 Number (%) of Patients LOVAZA label with TEAE Lovaza AMR101AMR101 4 g Placebo Placebo 2 g/day 4 g/day N = 226 N = 228 N = 76 N = 76N = 77 Event % % n (%) n (%) n (%) Subjects with 35.4 27.6 29 (38.2) 27(35.5) 28 (36.4) any TEAE Diarrhea 5 (6.6) 4 (5.3) 1 (1.3) Nausea 4(5.3) 5 (6.6) 1 (1.3) Dyspepsia  7 (3.1) 6 (2.6) 0 (0.0) 1 (1.3) 1 (1.3)Eructation 11 (4.9) 5 (2.2) 3 (3.9) 1 (1.3) 0 (0.0) Skin rash  4 (1.8) 1(0.4) 1 (1.3) 1 (1.3) 0 (0.0) Infection 10 (4.4) 5 (2.2)  9 (11.8) 2(2.6) 4 (5.2) Hypertension 0 (0.0) 4 (5.3) 1 (1.3) TEAE =treatment-emergent adverse event

1. A method of preventing stroke or myocardial infarction in a subjectin need thereof, comprising: providing a subject who is allergic to orhypersensitive to fish or seafood, is on statin therapy and has abaseline triglyceride level of at least 150 mg/dl; and administeringdaily to the subject a pharmaceutical composition comprising about 4 gof ethyl eicosapentaenoate, wherein the pharmaceutical composition issubstantially free of fish protein and is encapsulated in one or morecapsules.
 2. The method of claim 1 wherein the subject has or is at riskfor developing a cardiovascular-related disease.
 3. The method of claim2 wherein the subject has coronary heart disease or hypertension.
 4. Themethod of claim 3 wherein the subject does not develop skin rash afteradministration of the pharmaceutical composition.
 5. A method oftreating hypertriglyceridemia in a subject in need thereof, comprising:(a) providing a subject who has a fasting baseline triglyceride level ofat least about 500 mg/dl and who is allergic to fish or seafood, and (b)administering daily to the subject a pharmaceutical compositioncomprising about 4 g of ethyl eicosapentaenoate, wherein thepharmaceutical composition is substantially free of fish protein and isencapsulated in one or more capsules.
 6. The method of claim 5 whereinthe subject has or is at risk for developing a cardiovascular-relateddisease.
 7. The method of claim 6 wherein the subject has coronary heartdisease or hypertension.
 8. The method of claim 7 wherein the subjectdoes not develop skin rash after administration of the pharmaceuticalcomposition.